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Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3′-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation.
1. HOW DID OUR ANTIBODY PERFORM? CUSTOMER REVIEW: GAPDH Polyclonal Antibody (STJ96417)
2. WHAT WAS USED? Primary antibody: STJ96417. GAPDH Polyclonal Antibody. 100ul Provider: St John’s Laboratory Dilution ratio: 1:1000 Application: Western Blot. Image: In HeLa and Jurkat cell line, there were target bands detected by STJ96417 GAPDH Polyclonal Antibody.
3. WHAT WAS THE PROTOCOL? Treatment of materials: When cells were cultured to 70%-80% plating density, lysis buffer with protease inhibitors (Sigma) and equal volume of 2X loading buffer were added for 10 min water bath at 100?C. Gel electrophoresis: 10% polyacrylamide gel, constant voltage 160 V, 60 min. Transfer: 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min. Blocking: 1X TBST with 5% skimmed milk powder for 2 hr. Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder, 4?C for overnight. Membrane wash: 1X TBST wash for 3 times. Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% skimmed milk powder with 1:10000 dilution ratio, for 1.5 hr. Membrane wash: 1X TBST wash for 3 times. Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that the visualization time was identical for the same antigen).
4. WHAT WAS THE PROTOCOL? Gel electrophoresis information£º10% polyacrylamide gel, constant voltage 160 V, 60 min. Transfer information£º0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min. No. Antigen Loading amount Primary antibody Primary antibody dilution ratio Secondary antibody dilution ratio Target band KD Visualization time 1 HeLa lysate 10 ug STJ96417. GAPDH Polyclonal Antibody 1:1000 1:10000 36KD 130S 2 Jurkat lysate 10 ug 1:10000 36KD 130S
5. WHAT DID THE CUSTOMER THINK? Testing results were provided by BPI laboratory. They stated that ¡°In HeLa and Jurkat cell line, there were target bands detected by STJ96417 GAPDH Polyclonal Antibody¡±.
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