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Bradford protein assay
About: Bradford protein assay is a research topic. Over the lifetime, 635 publications have been published within this topic receiving 239107 citations.
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TL;DR: Abnormal yellow seminal plasma, compared to normal white seminal Plasma, had elevated levels of total protein and cholesterol and increased activities of acid phosphatase and aspartate aminotransaminase.
Abstract: Turkeys which produce yellow semen have abnormal ductuli efferentes’ epithelial morphology, with b!ebbing of cytoplasmic material into the ductal lumen. This could possibly increase the activity or concentration of seminal plasma components. In the present study, seminal plasma from 270 Large White breeder turkeys was evaluated for protein and cholesterol concentrations and the activities of acid phosphatase and asparate aminotransaminase. In a separate experiment, protein concentrations of turkey seminal plasma were estimated by biuret or Bradford methods. Bradford estimates were 46.6% less than those obtained with the biuret assay, using bovine serum albumin as the standard. Estimates of seminal plasma protein concentration in the main study were obtained using the Bradford method, and should be adjusted accordingly when compared with other studies using the biuret technique. Abnormal yellow seminal plasma, compared to normal white seminal plasma, had elevated levels of total protein and cholesterol and increased activities of acid phosphatase and aspartate aminotransasninase. Overall means were: 14.3 mg/mI, 38.9 mg/dl, 232.6 lU/mI, 81.0 lU/mI, respectively. Correlation coefficients for cholesterol concentration, acid phosphatase and aminotransaminase activity with protein concentration were +0.65, 0.70 and 0.50 (P<0.0001), respectively. Specific activities of both enzymes showed a significant reduction as seminal plasma protein increased, indicating a disproportionate increase in proteins other than these enzymes in yellow seminal plasma.
14 citations
TL;DR: In this article, phase pure (HAp) and fluoride-substituted apatite powders were synthesized by the hydrothermal method and their specific surface area was determined by atomic force microscopy (AFM).
14 citations
TL;DR: Substantial interference from N-hydroxysuccinimide (NHS) in the bicinchoninic acid (BCA) protein assay is reported, which indicates a similar BCA reaction mechanism for NHS and protein.
14 citations
Patent•
17 Dec 1984
TL;DR: In this article, the capacity of protein to bind silver was measured based on the optical density measured at 420 nm, which represents a 100-fold increase in sensitivity over the Coomassie Brilliant Blue dye binding procedure.
Abstract: A novel, inexpensive, highly sensitive and quantitative assay for measuring protein in solution based on the capacity of protein to bind silver is described. In this procedure, protein samples are first treated with glutaraldehyde and then exposed to ammoniacal silver. After a specified time, the reaction is terminated by the addition of sodium thiosulfate and the optical density measured at 420 nm. The useful range of the assay for the majority of standard proteins tested lies between 15 and 2000 ng. This represents a 100-fold increase in sensitivity over the Coomassie Brilliant Blue dye binding procedure. There is little or no interference from carbohydrates, non-ionic detergents or ethanol. Pretreatment of protein samples with Bio-Gel P-2 to remove salts, thiol agents, EDTA and SDS, makes this procedure compatible with most commonly used buffers.
14 citations
TL;DR: This assay appears to specifically measure total sPD-1 protein since the human anti- PD-1 antibody, nivolumab, and the endogenous ligands of PD- 1 protein, PDL-1 and PDL,2, do not interfere with the assay.
Abstract: Programmed death-1 (PD-1) protein is a co-inhibitory receptor which negatively regulates immune cell activation and permits tumors to evade normal immune defense. Anti-PD-1 antibodies have been shown to restore immune cell activation and effector function—an exciting breakthrough in cancer immunotherapy. Recent reports have documented a soluble form of PD-1 (sPD-1) in the circulation of normal and disease state individuals. A clinical assay to quantify sPD-1 would contribute to the understanding of sPD-1-function and facilitate the development of anti-PD-1 drugs. Here, we report the development and validation of a sPD-1 protein assay. The assay validation followed the framework for full validation of a biotherapeutic pharmacokinetic assay. A purified recombinant human PD-1 protein was characterized extensively and was identified as the assay reference material which mimics the endogenous analyte in structure and function. The lower limit of quantitation (LLOQ) was determined to be 100 pg/mL, with a dynamic range spanning three logs to 10,000 pg/mL. The intra- and inter-assay imprecision were ≤15%, and the assay bias (percent deviation) was ≤10%. Potential matrix effects were investigated in sera from both normal healthy volunteers and selected cancer patients. Bulk-prepared frozen standards and pre-coated Streptavidin plates were used in the assay to ensure consistency in assay performance over time. This assay appears to specifically measure total sPD-1 protein since the human anti-PD-1 antibody, nivolumab, and the endogenous ligands of PD-1 protein, PDL-1 and PDL-2, do not interfere with the assay.
14 citations