(MS1078)
GSK3 beta Total + pSer9 Human Flow Cytometry Assay Kit (ab131381)
Overview
- Product nameGSK3 beta Total + pSer9 Human Flow Cytometry Assay Kit
- Tests1 x 96 test
- Sample typeAdherent cells, Suspension cells
- Assay typeDirect
- Species reactivityReacts with: Human
- Product overview
ab131381 is a panel of antibodies that measure the protein levels of GSK3B total and phosphorylation of GSK3B at serine residue 9. The assay combines the power of single cell analysis obtained with flow cytometry and the specificity of antibody-based immunostaining to quantitate protein and phosphorylation levels in cultured cells. Cells are harvested and fixed/permeabilized in suspension, targets of interest are detected indirectly with highly specific, well-characterized monoclonal antibodies that are then labeled with fluorescent antibodies.
This flow cytometry panel generates quantitative data with specificity similar to Western blotting, but with much greater quantitative precision. This method rapidly fixes the cells in-situ, stabilizing the in-vivo levels of proteins, and thus essentially eliminates changes during sample handling, such as preparation of protein extracts.Glycogen synthase kinase 3 is a proline directed serine, threonine kinase originally identified due to its ability to phosphorylate and inactivate glycogen synthase and later found to be of key importance in signaling pathways, cell fate determination, energy metabolism, transcription regulation, neuronal development and body pattern formation. There are two isoforms of GSK-3 (alpha and beta) which show a high degree of homology within their catalytic domains.
- Tested applicationsFlow Cyt more details
Properties
- Storage instructionsStore at +4°C. Please refer to protocols.
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Components 1 x 96 tests 100X Triton X-100 1 x 1.25ml 10X Phosphate Buffered Saline 1 x 100ml 50X Antibody Cocktail Mouse (Anti-GSK3B total + Rabbit Anti-GSK3B pS9 Antibody) 1 x 220µl Blocking Solution 1 x 10ml - Research Areas
Applications
Our Abpromise guarantee covers the use of ab131381 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| Flow Cyt | Flow Cyt |
GSK3 beta Total + pSer9 Human Flow Cytometry Assay Kit images
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Flow Cytometry - GSK3 beta Total + pSer9 Human Flow Cytometry Assay Kit (ab131381)Antibody specificity demonstrated by Flow cytometry. Non-induced Jurkat cells (red) and calyculin induced (blue) were targeted with the antibody cocktail against GSK3B total and GSK3B (pSer9). Background fluorescence (pink) was determined with a no-primary antibody control. Primary antibodies were labeled with 1:500 dilution of GAM IgG – H&L DyLight® 488 Cat# ab96879 and 1:500 dilution of GAR IgG – H&L DyLight® 649 Cat# ab96902 respectively. Signal intensity was measured in FL-1 (GSK3B total) and FL-4 (GSK3B pSer9). After background subtraction, the Calyculin induced cell line shows a 6 fold increase in the levels of phosphorylated GSK3B in comparison to a 1.2 decrease in the levels of GSK3B protein. -
Immunocytochemistry/ Immunofluorescence - GSK3 beta Total + pSer9 Human Flow Cytometry Assay Kit (ab131381)Antibody specificity demonstrated by immunocytochemistry. ICC was carried out on HeLa cells treated with Calyculin (top) or vehicle (bottom) with Rabbit anti-GSK3B pSer9 and Mouse anti-GSK3B using all buffer reagents as supplied in this kit. Labeling was carried out with 1:500 dilution of GAR IgG - H&L DyLight® 594 Cat# ab96897 and 1:500 dilution of GAM IgG – H&L DyLight® 488 Cat# ab96879 respectively. The calyculin induced cells (top) show a significant induction of GSK3B phosphorylation at residue S9 in comparison to the non-induced control (bottom). -
Western blot - GSK3 beta Total + pSer9 Human Flow Cytometry Assay Kit (ab131381)All lanes : GSK3 beta Total + pSer9 Human Flow Cytometry Assay Kit (ab131381)
Lane 1 : GSK3B protein (ab63193) at 0.05 µg
Lane 2 : Jurkat cells treated with DMSO at 40 µg
Lane 3 : Jurkat cells treated with 100 nM Calyculin at 40 µg
Lane 4 : HeLa cells treated with DMSO at 40 µg
Lane 5 : HeLa cells treated with 100 nM Calyculin at 40 µg
Protocols
References for GSK3 beta Total + pSer9 Human Flow Cytometry Assay Kit (ab131381)
ab131381 has not yet been referenced specifically in any publications.
