Why is the pH differ between separating gel and stacking gel in SDS-PAGE?
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The pH difference between the separating gel and stacking gel in SDS-PAGE is due to the different buffer compositions used in each gel. The stacking gel uses the principles of isotachophoresis to concentrate samples into small zones without separating them . It includes excess SDS and a thiol reagent, which denatures the protein mixture and disrupts disulfide bonds. Under these conditions, all reduced polypeptides bind the same amount of SDS, independent of amino acid composition and sequence. The resolving power of SDS-PAGE is greatly enhanced by the inclusion of a stacking gel. On the other hand, the separating gel is designed to separate the negatively charged SDS-protein complexes based on molecular weight differences . Therefore, the pH difference between the separating gel and stacking gel is necessary to create the appropriate conditions for both concentration and separation of the protein samples in SDS-PAGE.
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| Papers (5) | Insight |
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15 Citations | The provided paper does not provide information about the pH difference between the separating gel and stacking gel in SDS-PAGE. |
16 Citations | The provided paper is about sample stacking during membrane-mediated loading in automated DNA sequencing. It does not provide information about the pH difference between separating gel and stacking gel in SDS-PAGE. |
| The provided paper is about capillary zone electrophoresis (CZE), not SDS-PAGE. Therefore, the paper does not provide information about the pH difference between separating gel and stacking gel in SDS-PAGE. | |
| The provided paper does not mention the pH difference between the separating gel and stacking gel in SDS-PAGE. | |
| The provided paper is about a novel polyacrylamide gel electrophoresis (PAGE) method for stacking and separating human hemoglobins. It does not mention SDS-PAGE or the pH difference between separating gel and stacking gel. |
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