Customer Review For GSK3beta Polyclonal Antibody (STJ93447)


Target information

GSK3beta Polyclonal Antibody is an active protein kinase that acts as a negative regulator in the hormonal control of glucose homeostasis, Wnt signaling and regulation of transcription factors and microtubules, by phosphorylating and inactivating glycogen synthase (GYS1 or GYS2), EIF2B, CTNNB1/beta-catenin, APC, AXIN1, DPYSL2/CRMP2, JUN, NFATC1/NFATC, MAPT/TAU and MACF1. Requires primed phosphorylation of the majority of its substrates.

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1. HOW DID OUR ANTIBODY PERFORM? CUSTOMER REVIEW: GSK3beta Polyclonal Antibody (STJ93447)
2. WHAT WAS USED? Primary antibody: STJ93447. GSK3beta Polyclonal Antibody. 100ul Provider: St John’s Laboratory Dilution ratio: 1:1000 Application: Western Blot. Image: In HeLa cell line, the target band was detected by STJ93447. GSK3beta Polyclonal Antibody. In A549 cell line, there was no target band detected by STJ93447 GSK3beta 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 St J93447. GSK3beta Polyclonal Antibody 1:1000 1:10000 47KD 30S 2 A549 lysate 10 ug 1:10000 47KD 30S
5. WHAT DID THE CUSTOMER THINK? Testing results were provided by BPI laboratory. They stated that ¡°In HeLa cell line, the target band was detected by GSK3beta Polyclonal Antibody (STJ93447). In A549 cell line, there was no target band detected by GSK3beta Polyclonal Antibody (STJ93447).¡±
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