Showing papers on "Bradford protein assay published in 2016"

Formulation and optimization of oxaliplatin immuno-nanoparticles using Box–Behnken design and cytotoxicity assessment for synergistic and receptor-mediated targeting in the treatment of colorectal cancer

Abstract: Conventional chemotherapy majorly lacks clinical application attributed to its inspecificity, adverse effects and inability to penetrate into tumor cells. Hence, the aim of the study was to prepare oxaliplatin solid lipid nanoparticles (OP-SLN) by microemulsion method optimizing it by Box-Behnken design and then covalently conjugated to TRAIL (CD-253) monoclonal antibody (TR-OP-SLN) for targeting colorectal cancer cells. The optimized OP-SLN3 has shown an appreciable particle size (121 ± 1.22 nm), entrapment efficiency (78 ± 0.09%) and drug loading (32 ± 1.01%). Fluorescence study and the Bradford assay further confirmed the binding of the protein. A 1.5-fold increase in cytotoxicity of immuno-nanoparticles (4.9 μM) was observed.

Smartphone for Point-of-Care Quantification of Protein by Bradford Assay

Abstract: We address in this paper a powerful point-of-care platform to conduct the Bradford assay. Our method was based on smartphone for colorimetric quantification of total protein in human blood plasma, presenting low cost, simplicity, portability, autonomy and ability for remote transmission of the data. Other advantage concerns the high number of smartphone's users worldwide. This feature contributes for the application of the method by non-specialist people. The interferences from external light were successfully solved by illuminating the samples with a straightforward negatoscope. Our method generated a satisfactory exactness, with accuracy percentages ranging from 87.2 to 99.1%.

Comparable efficiency of different extraction protocols for wheat and rye prolamins

Abstract: The identification and quantification of cereal storage proteins is of interest of many researchers. Their structural or functional properties are usually affected by the way how they are extracted. The efficiency of extraction process depends on the cereal source and working conditions. Here, we described various commonly used extraction protocols differing in the extraction conditions (pre-extraction of albumins/globulins, sequential extraction of individual protein fractions or co-extraction of gluten proteins, heating or non-heating, reducing or non-reducing conditions). The total protein content of all fractions extracted from commercially available wheat and rye flours was measured by the Bradford method. Tris-Tricine SDS-PAGE was used to determine the molecular weights of wheat gliadins, rye secalins and high-molecular weight glutelins which are the main triggering factors causing celiac disease. Moreover, we were able to distinguish individual subunits (α/β-, γ-, ω-gliadins and 40k-γ-, 75k-γ-, ω-secalins) of wheat/rye prolamins. Generally, modified extraction protocols against classical Osborne procedure were more effective and yields higher protein content in all protein fractions. Bradford measurement led into underestimation of results in three extraction procedures, while all protein fractions were clearly identified on SDS-PAGE gels. Co-extraction of gluten proteins resulted in appearance of both, low-molecular weight fractions (wheat gliadins and rye secalins) as well as high-molecular weight glutelins which means that is not necessary to extract gluten proteins separately. The two of three extraction protocols showed high technical reproducibility with coefficient of variation less than 20%. Carefully optimized extraction protocol can be advantageous for further analyses of cereal prolamins.

A Dynamic Interaction of Coomassie Dye with the Glycine Transporters N-termini.

Abstract: Coomassie Brilliant Blue interacts with proteins and even though the interactions exhibit variation due to the amino acid content, reported dye interactions with individual proteins appear to be relatively stable. Here we report an atypical dynamic interaction of glycine transporters 1 and 2 N-termini with Coomassie dye, resulting in intramolecular interference with their Bradford assay. These proteins exhibit classic protein-Coomassie G-250 complex with absorption maximum at 595 nm, which within minutes starts to decrease and parallel increase of absorbance shoulders above 300 and 700 nm is observed. Interestingly, these effects are eliminated upon fusion of glycine transporters N-termini with glutathione S-transferase protein or by the presence of glutathione S-transferase or bovine serum albumin in the same solution. Circular dichroism data revealed largely unstructured character of glycine transporters N-termini, which suggests that dynamic properties of these protein- Coomassie complexes might be a signature of high flexibility and protein disorder. The assay might potentially reveal similar domains in other proteins and help to associate them with particular functions.

COMPARISON STUDY OF THERAPEUTIC PROPERTIES OF PROTEINS AND SECONDARY METABOLITES FROM CARICA PAPAYA Original Article

Abstract: Objective: The current study aims to compare the therapeutic potential of protein extracts and secondary metabolites extracts using methanol, ethyl acetate and hexane from both the fruits and seeds of Carica papaya. Methods: All of the crude proteins and secondary metabolite fractions extracted from the fruits and seeds of Carica papaya were assessed and compared in DPPH free-radical scavenging assay for antioxidant activities and brine shrimp lethality assay for cytotoxic potentials. Protein content was quantified by Bradford assay while total phenolic and flavonoids contents were determined by Folin-Ciocalteu and aluminum chloride colorimetric methods accordingly. Bioactive protein molecules and secondary metabolite fractions were then characterized and analyzed in SDS- PAGE and silica thin layer chromatography respectively. Results: Evidence demonstrated that the secondary metabolite extracts of ethyl acetate fraction of the seeds of Carica papaya have the highest antioxidant activity (IC 50 value of 25.97 µg/ml) as well as cytotoxicity activity (LC 50 value of 142.27 µg/ml) in comparison to other crude proteins, methanol or hexane extracts. Notably, crude protein from fruits possesses relatively high antioxidant (IC 50 value of 34.62 µg/ml) and cytotoxicity (LC 50 Conclusion: This study proved that the crude protein from fruits and ethyl acetate fraction from seeds and fruits of Carica papaya have great potential to be further developed into therapeutic drugs in future. value of 222.52 µg/ml) activities. The presence of bioactive phenolic and flavonoid in crude secondary metabolite extracts and bioactive protein molecules in crude protein extracts confer the high antioxidant capacity and cytotoxic potential of Carica papaya.

Comparison study of therapeutic properties of proteins and secondary metabolites from carica papaya

Abstract: Objective: The current study aims to compare the therapeutic potential of protein extracts and secondary metabolites extracts using methanol, ethyl acetate and hexane from both the fruits and seeds of Carica papaya . Methods: All of the crude proteins and secondary metabolite fractions extracted from the fruits and seeds of Carica papaya were assessed and compared in DPPH free-radical scavenging assay for antioxidant activities and brine shrimp lethality assay for cytotoxic potentials. Protein content was quantified by Bradford assay while total phenolic and flavonoids contents were determined by Folin-Ciocalteu and aluminum chloride colorimetric methods accordingly. Bioactive protein molecules and secondary metabolite fractions were then characterized and analyzed in SDS-PAGE and silica thin layer chromatography respectively. Results: Evidence demonstrated that the secondary metabolite extracts of ethyl acetate fraction of the seeds of Carica papaya have the highest antioxidant activity (IC 50 value of 25.97 µg/ml) as well as cytotoxicity activity (LC 50 value of 142.27 µg/ml) in comparison to other crude proteins, methanol or hexane extracts. Notably, crude protein from fruits possesses relatively high antioxidant (IC 50 value of 34.62 µg/ml) and cytotoxicity (LC 50 value of 222.52 µg/ml) activities. The presence of bioactive phenolic and flavonoid in crude secondary metabolite extracts and bioactive protein molecules in crude protein extracts confer the high antioxidant capacity and cytotoxic potential of Carica papaya . Conclusion: This study proved that the crude protein from fruits and ethyl acetate fraction from seeds and fruits of Carica papaya have great potential to be further developed into therapeutic drugs in future. Keywords: Carica papaya , Antioxidant, Cytotoxicity, Crude fruit protein, Phenolic, Flavonoid

Method for detecting differential protein produced by action between alpinetin or cardamonin and serum

Abstract: The invention provides a method for detecting differential protein produced by action between alpinetin or cardamonin and serum. The method comprises the following steps: (1) shaking up serum protein and alpinetin or cardamonin with the concentration of 1.0*10 mol/L-3.0*10 mol/L and cultivating at the constant temperature; (2) removing highly-abundant proteins according to the steps of a serum albumin scavenger; (3) quantifying an obtained protein sample by use of the Bradford method; (4) performing isoelectric focusing according to the protein loading quantity of 1300 [mu]g/24 cm of a rubber strip; (5) transferring for polyacrylamide gel electrophoresis after balancing of the rubber strip; (6) after finish of electrophoresis, performing Coomassie brilliant blue staining, decolorization, storage and scanning analysis; (7) performing enzymolysis and mass spectrum identification on differential protein spots.

[Spectrophotometric determination of protein content in THP-1 monocytes/macrophages - description of the method].

Abstract: Proteins are the basic building block of tissue, and are part of enzymes and hormones regulating many important life processes. Changes in their concentration control the metabolic processes of the cell. Quantitative determination of the protein content is divided into indirect methods (e.g. Kjeldahl method) and direct methods (buret method, Lowry, immunoenzymatic, formol method, based on incorporation of dye in the range of ultraviolet spectrophotometry, and based on the phenomenon of selective absorption of radiation in the infrared range). One of the methods for the determination of protein content is the spectrophotometric method described by Bradford. The protein concentration assay procedure utilizes the phenomenon of formation of the dye (Coomassie Brillant Blue G-250)-protein and colour intensity is proportional to the protein content in the solution. The aim of this study was to verify the usefulness of this method for determining the protein content in THP-1 cells cultured with extracts of nettle fruit stalks (Urtica dioica L.). Aqueous and alcohol extracts at two concentrations were used. It has been shown that the spectrophotometric determination of protein content by the Bradford method is an effective and accurate method for determining the concentration of protein in THP-1 macrophages. The results indicate that this method can be recommended for the determination of the protein content in other cell cultures.

The Use of Pulsed Electric Fields for Protein Extraction from Nanochloropsis and Chlorella

Abstract: Classical methods, used for large scale treatments such as mechanical or chemical extractions, affect the integrity of extracted cytosolic protein by releasing proteases contained in vacuoles. Our previous experiments on batch and flow processes electro-extraction on yeasts proved that pulsed electric field technology was highly effective and preserve the activity of released cytosolic proteins. As induction of electro-permeabilization is under the control of target vesicle size, protocols were chosen to affect only the plasma membrane and not vacuole membranes. Furthermore, large cell culture volumes are easily treated by the flow technology. Based on this previous knowledge, we developed a new protocol in order to electro-extract total cytoplasmic proteins from microalgae (Nannochloropsis salina, Chlorella vulgaris). Due to the size selective effect of electro-permeabilization, as the mean diameter for N. salina is only 2.5 μm, we used stronger field strengths than for C. Vulgaris. A train of repetitive 2 ms long pulses of alternating polarities was delivered. The electric treatment was followed by a 24 h incubation period in a salty buffer containing DTT at room temperature. The amount of total proteins release was observed by a classical Bradford assay. A more accurate evaluation of protein release was obtained by SDS-PAGE. Similar results were obtained with C. vulgaris and N. Salina under optimized electrical conditions.

Optimization of amino acid-stabilized erythropoietin parenteral formulation: In vitro and in vivo assessment.

Abstract: The aim of this study was to optimize the formulation of erythropoietin (EPO) using amino acids instead of human serum albumin (HSA) and to evaluate its in vivo stability in order to avoid the risk of viral contamination and antigenicity. Different EPO formulations were developed in such a way as to allow studying the effects of amino acids and surfactants on the EPO stability profile. The main techniques applied for EPO analysis were ELISA, Bradford method, and SDS gel electrophoresis. The in vivo stability was evaluated in a Balb-c mouse animal model. The results showed that the presence of surfactant was very useful in preventing the initial adsorption of EPO on the walls of vials and in minimizing protein aggregation. Amino acid combinations, glycine with glutamic acid, provided maximum stability. Formulation F4 (containing glycine, glutamic acid and Tween 20) showed minimum aggregation and degradation and in vivo activity equivalent to commercially available HSA-stabilized EPO (Eprex®).

Whole-course C-reactive protein assay kit and application thereof

Abstract: The invention discloses a whole-course C-reactive protein assay kit and an application thereof. The kit includes the following components: an M reagent, an R1 reagent, an R2 reagent, an alkaline pre-treatment solution, a pre-excitation agent and an excitation agent. The M reagent includes 0.5-1 mg/ml of magnetic particles coated by a first antibody, 0.5-1% of casein and 0.5-1% of bovine serum albumin, wherein the solvent is a phosphate buffer solution being 7.0-8.0 in pH. The coating quantity of the first antibody is 5-15 [mu]g per mg of the magnetic particles. The R1 reagent is an HCl solution being 10-20 mM in concentration. The R2 reagent includes acridinium ester which is coated by a second antibody, 0.5-1% of casein and 0.5-1% of bovine serum albumin, wherein the solvent is a phosphate buffer solution being 7.0-8.0 in pH. The coating quantity of the second antibody is 5-15 [mu]g per ml of the acridinium ester. The alkaline pre-treatment solution is a NaOH solution being 20-50 mM in concentration. Strong alkali is added to the kit during reaction to damage the proteins, and then strong acid is added to neutralize the reaction system. The kit is 0.02-100 mg/l in detection range, and the reagent can reach the requirement of detection of the whole course of C-reactive protein.

Diffusion Behavior of Microbial Transglutaminase to Induce Protein Crosslinking in Oil-in-Water Emulsions

Abstract: Food-grade hydrogel particles composed of sodium alginate were used to investigate the diffusion behavior of microbial transglutaminase (mTG) to induce crosslinking of interfacially adsorbed protein. For this purpose, mTG-loaded hydrogel beads were mixed with caseinate-stabilized oil-in-water emulsions, whereas Bradford assay and ammonia measurements were utilized to monitor the enzyme-induced interfacial protein crosslinking. Different alginate (0.5–1.5%) and gelling concentrations (50–500 mM CaCl2) were used to modulate the hydrogel mesh size and number of junction zones. The results indicated that mTG was able to diffuse out of alginate beads. However, a decrease in NH3 concentration with increasing alginate and CaCl2 levels was observed due to the formation of tight and dense bead structures. These results illustrate that the spatial distribution of molecules in complex matrices plays a key role on the enzyme accessibility

Method for acquiring two-dimensional electrophoretogram of nectar protein of liriodendron tulipifera by aid of two-dimensional electrophoresis system

Abstract: The invention discloses a method for acquiring a two-dimensional electrophoretogram of nectar protein of liriodendron tulipifera by the aid of a two-dimensional electrophoresis system. After a nectar sample is filtered by an ultramicro filter membrane, the nectar protein is concentrated and purified by a protein ultrafiltration and concentration purification column, the concentration of the purified and concentrated nectar protein is detected with a Bradford method, the molecular weight distribution range of the nectar protein of the liriodendron tulipifera is pre-detected through one-dimensional lauryl sodium sulfate polyacrylamide gel electrophoresis, finally, proper standard protein and a gel strip with a proper length and pH range are selected according to a one-dimensional electrophoresis result, the nectar protein of the liriodendron tulipifera is separated with the two-dimensional electrophoresis system, and the two-dimensional electrophoretogram is acquired. The method has the characteristics that the operation is simple, the protein extraction rate is high and few interfering substances exist and is suitable for special materials such as liquid samples, the high-quality two-dimensional gel electrophoretogram with high resolution ratio, clear protein points, larger numbers, uniform distribution and clear background can be obtained, and the experiment repeatability and the stability are good.

A single step protein assay that is both detergent and reducer compatible: The cydex blue assay

Abstract: Determination of protein concentration in often an absolute pre-requisite in preparing samples for biochemical and proteomic analyses. However, current protein assay methods are not compatible with both reducers and detergents, which are however present simultaneously in most denaturing extraction buffers used in proteomics and electrophoresis, and in particular in SDS electrophoresis. We found that inclusion of cyclodextrins in a Coomassie blue-based assay made it compatible with detergents, as cyclodextrins complex detergents in a 1:1 molecular ratio. As this type of assay is intrinsically resistant to reducers, we have thus developed a single step assay that is both detergent and reducer compatible. Depending on the type and concentration of detergents present in the sample buffer, either beta-cyclodextrin or alpha-cyclodextrin can be used, the former being able to complex a wider range of detergents and the latter being able to complex higher amounts of detergents due to its greater solubility in water. Cyclodextrins are used at final concentrations of 2-10 mg/mL in the assay mix. This typically allows to measure samples containing as little as 0.1 mg/mL protein, in the presence of up to 2% detergent and reducers such as 5 % mercaptoethanol or 50 mM DTT in a single step with a simple spectrophotometric assay. This article is protected by copyright. All rights reserved.

A single step protein assay that is both detergent and reducer compatible: The cydex blue assay

Abstract: Determination of protein concentration in often an absolute pre-requisite in preparing samples for biochemical and proteomic analyses. However, current protein assay methods are not compatible with both reducers and detergents, which are however present simultaneously in most denaturing extraction buffers used in proteomics and electrophoresis, and in particular in SDS electrophoresis. We found that inclusion of cyclodextrins in a Coomassie blue-based assay made it compatible with detergents, as cyclodextrins complex detergents in a 1:1 molecular ratio. As this type of assay is intrinsically resistant to reducers, we have thus developed a single step assay that is both detergent and reducer compatible. Depending on the type and concentration of detergents present in the sample buffer, either beta-cyclodextrin or alpha-cyclodextrin can be used, the former being able to complex a wider range of detergents and the latter being able to complex higher amounts of detergents due to its greater solubility in water. Cyclodextrins are used at final concentrations of 2-10 mg/mL in the assay mix. This typically allows to measure samples containing as little as 0.1 mg/mL protein, in the presence of up to 2% detergent and reducers such as 5 % mercaptoethanol or 50 mM DTT in a single step with a simple spectrophotometric assay. This article is protected by copyright. All rights reserved.

Fractionation and identification of the allergic proteins in Aspergillus species

Abstract: Background and purpose Allergy is an undesired immune response to non-pathogenic agents. However, some opportunistic microorganisms such as fungi can also cause allergy. Among those fungi, hyphae form of Aspergillus strains including A. fumigatus, A. flavus, and A. niger could be mentioned. In this study, we aimed to separate allergic proteins from Aspergillus strains and determine their identity. Materials and methods Standard species of Aspergillus strains were cultivated in optimized conditions and the mycelium was separated by centrifugation. The fungal cells were lysed through physical methods such as freeze-thawing and grinding to prepare a suitable protein extract. The protein concentration was measured by Bradford method and the electrophoretic pattern of the extract was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were fractionated by ammonium sulfate precipitation and anion exchange chromatography using fast protein liquid chromatography (FPLC) system. The IgE immunoreactivity of the sensitized patients and controls was studied using the fractionated proteins by enzyme-linked immunosorbent assay (ELISA). Following SDS-PAGE, proteins were electrotransferred onto polyvinylidene difluoride (PVDF) membranes and the strips were blotted with allergic patients' and controls' sera. The immunoreactive bands were excised from colloidal coomassie-stained SDS-PAGE gels and studied by mass spectroscopy methods. Results Among the studied species, A. fumigatus showed stronger IgE reactivity and more IgE reactive protein bands than others did. The proteins with higher molecular weights showed stronger immunoreactivity in Western blotting. Receiver operating characteristic curve analysis demonstrated a correlation between the results of the applied ELISA methods. One of the most prominent IgE-reactive proteins was confirmed to be 45 kDa mycelia catalase. Conclusion Our findings confirmed that high molecular weight proteins might play a major role in allergy and IgE reactivity to Aspergillus species. Moreover, the results showed that precipitation and chromatographic methods are applicable for fractionation of fungal proteins such as mycelial catalase.

Studies on the Site-specific PEGylation Induced Interferences Instigated in Uricase Quantification Using the Bradford Method

Abstract: Uricase from Bacillus fastidiosus was site-specifically PEGylated using methoxypolyethyleneglycol-maleimide (mPEG-mal) of different molecular weights (750 Da, 5 kDa, 10 kDa) via Thiol PEGylation strategy. The obtained monoPEGylated uricase conjugates were characterized using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and the molecular weight of single subunit of the conjugate was found to be 42.6, 48.1 and 56.3 kDa with respect to different molecular weights of m-PEG-mal. The influence of PEGylation on the quantification of uricase using protein quantification techniques like Bradford assay, RP-HPLC detection and Dumbroff method has been evaluated. A gradual decline in the absorbance value was observed with the advancement of the PEGylation reaction, indicating an interferences in the protein quantification due to PEGylation. The extent of interference highly dependence on mPEG-mal concentration and its chain length. The present study indicates that the quantification of PEGylation induced interferences caused in protein measured ought to be prudently measured at every discrete step of the PEGylation process.

Evaluation of renal epithelial cell protein under stress condition

Abstract: Objective: Proteins are an important component of cells which are involved in various cellular functions. Different kind of stressing conditions has different responses in the components of the protein synthesis system. Super saturation condition in kidney environment leads to crystallization process. Crystals thus formed injure the surrounding cells and result in reactive oxygen species (ROS) formation. There might be some changes in the protein synthesis when the kidney cells enter in oxidative stress. In the present study, kidney cell lines were exposed to oxidative stress and their proteins were analyzed using Bradford analysis and SDS-PAGE. Methods: Vero cells were obtained from NCCS Pune and cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) and maintained in a humidified incubator at 37 °C with 5% CO 2. Calcium phosphate (CaP) crystals were prepared by the homogeneous system. After FTIR analysis crystals were used to injure Vero cell line. H 2 O 2 was also used to injure the Vero cells. Intracellular protein was extracted from healthy cells and injured cells (with CaP crystals and H 2 O 2 ). Ammonium sulfate precipitation method was used for the isolation of extracellular protein from the media of healthy and injured cells. Bradford method was used for the quantitative estimation of protein. Extracted proteins were analyzed by SDS-PAGE. Results: Amount of intracellular and extracellular protein of normal cells was 4.84±0.004µg/ml. Intracellular protein of CaP injured and H 2 O 2 injured cells were 10.59±0.003 µg/ml and 10.78±0.011µg/ml respectively. While extracellular protein of injured cells was nearly 4 µg/ml. Intracellular protein bands ranging from 14.3 to 97.4 kDa was observed in healthy cells. Protein bands of ~40kDa and ~20kDa was absent in H 2 O 2 and CaP injured intracellular protein extract. Two extracellular protein bands of 66kDa and ~60kDa were present in injured cells and healthy cells. Conclusion: When exposed to oxidative stress several proteins are oxidized decreasing the activity of many metabolic pathways. In the present study amount of intracellular protein increases when cells are injured with CaP or H 2 O 2. While extracellular protein remains more or less same in both healthy and injured condition of cells. In SDS-PAGE analysis few bands were missing in the intracellular extract of injured cells. These results indicate that the amount of protein varies when cells are injured with CaP and H 2 O 2.

Expression and purification of recombinant Shiga toxin 2B from Escherichia coli O157:H7

Abstract: Enterohemorrhagic Escherichia coli are important human food-borne pathogens. Recently, Shiga toxin-producing E. coli (STEC) causes life-threatening hemolytic-uremic syndrome (HUS). In this study, Stx2B gene, a subunit of Shiga toxin, was amplified via polymerase chain reaction (PCR) from the chromosomal DNA of clinical fecal sample using appropriate primers. The PCR product was cloned to commercially available plasmid pH6HTN His6HaloTag® T7 containing two purification tags, namely, six histadine tag and Halo tag. The integrity of the constructed plasmid was confirmed using restriction enzyme mapping and sequencing. Then, Stx2B protein expressed after induction with isopropyl β-D-1-thiogalactopyranoside (IPTG) in E. coli JM109 (DE3) under the control of the T7 promotor. The two step purification trains were used to purify native Stx2B. First step purification was Ni-immobilized metal ion affinity chromatography (IMAC) column, followed by second step using HaloLink resin. The native Stx2B was obtained after column cleavage of halo-tag using HaloTEV protease. Maximum protein expression of Stx2B economically was obtained using 1 mM IPTG for 4 h at 37°C. Protein identity was confirmed by a band at ~11.4 kDa using 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and StxB2 yield was 450 µg ml-1 confirmed by Bradford assay. Recombinant Stx2B protein was produced in highly pure yield using HaloTag technology. Key words: Escherichia coli O157:H7, StxB gene, expression, HaloTag technology, purification.

Determination and analysis of protein profile of different transfer factors

Abstract: Background: The transfer factor (TF) is the dialyzable extract of leukocytes with cellular immunity transfer properties. Its use has spread in the treatment of a wide range of immunologic, infectious, and even oncological diseases. However, important aspects in their protein profile, component concentrations, and a well-defined action mechanism are not completely unknown. Objectives: To analyze the protein profiles of different transfer factors marketed in Mexico. Methods: 6 TF marketed in Mexico were obtained and analyzed, quantifying protein with thaze Bradford method, by high-performance liquid chromatography (HPLC), and polyacrylamide gel electrophoresis (SDS-PAGE). All samples were analyzed in duplicate. Results : The total protein concentrations of all TF analyzed are less than 0.2 mg/mL. The chromatographic profiles showed differences in some TF. The protein concentration was 6 to almost one thousand times lower compared to reports by some manufacturers. Conclusion: Almost all transfer factors marketed in Mexico lack a labeling and health record that meets the official standards.

Method for detecting angiogenesis promoting capacity of MSC through VCAM-1

Abstract: The invention discloses a method for detecting the angiogenesis promoting capacity of an MSC through VCAM-1. The method is characterized in that a new purpose of known protein VCAM-1 is found, the protein expression of VCAM-1 is detected through an ELISA, the total protein content is measured through a Bradford method, and the angiogenesis promoting capacity of the cell is reflected through the protein content of VCAM-1 in every 1 g of total protein. The angiogenesis promoting capacity of the MSC can be quantified through the method.

Estimation of protein concentration in cell lines after treated with toxin a extracted from pseudomonas aeruginosa

Abstract: Two cell lines REF (Primary Rat Embryo Fibroblast) and AMN3 (Ahmed-Mohammed-Nahi-2003) were used to investigated the role of toxin A extracted from pseudomonas aeruginosa in inhibition of protein synthesis in cell lines after exposure to purified toxin A. Results have shown that incubation of toxin A (purified toxin A alone in first group and purified toxin A activated with urea in the second group) at the concentration (5, 25, 50, 100) ng /ml with both REF and AMN3 cell lines for 48 hrs will cause significant decrease in protein concentration in both cell lines in comparison with untreated control when estimated by Bradford method.