WebMay 11, 2024 · Western blotting procedure to detect phosphorylated proteins. A) Proteins are separated via SDS-PAGE where lane 1 = protein ladder, lane 2 = unphosphorylated protein, and lane 3 = phosphorylated protein. ... The isotype control ensures that the MFI of the phospho-antibody is specific for the phosphorylated protein-of-interest rather than an ... WebPanprotein(700nm) Treatment Replicate Primary Antibody Signal Average StDev %CV - 1 pan 990 950 36 4% - 2 pan 940 - 3 pan 920 + 1 pan 930 943 51 5% + 2 pan 900
Western Blotting Protocol (Primary Ab Incubation In BSA)
WebWestern Blotting Protocol (Primary Ab Incubation In BSA) For Western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween-20 at 4°C with gentle shaking, overnight. Products available from Cell Signaling Technology are linked by their respective catalog numbers. A. Solutions and Reagents WebIncrease the blocking incubation period and consider changing the blocking agent. We recommend blocking 3–5% non-fat dry milk, BSA, or normal serum for 1 hr at room temperature. Incubation temperature may be too high. Make sure you incubate samples at 4°C. Keep on ice throughout the western blot process. crystal glass hours
Phospho-Akt (Ser473) Antibody Cell Signaling Technology
WebJul 11, 2024 · For alkaline phosphatase-based detection and Western blotting of phosphorylated proteins, use TBS-based buffers, but for most other Western blot applications, both PBS and TBS buffers are largely interchangeable. However, don’t be afraid to challenge your SOP -- you might find that using a different buffer works wonders. WebWESTERN BLOTTING OF PHOSPHO-PROTEINS PROTOCOL Homogenize the cells or tissue of interest in lysis buffer made fresh and containing a cocktail of protease inhibitors (and phosphatase inhibitors when dealing with phosphorylated proteins). As soon as lysis … WebAnti-phospho-Histone H2A.X (Ser139), clone 6L16 ZooMAb, is a recombinant Rabbit Monoclonal Antibody that specifically targets phospho Histone H2AX (ser139) and has been tested for use in Immunocytochemistry, Immunohistochemistry (Paraffin), Peptide Inhibition Assay, and Western Blotting. crystal glass inc muncie