Glycogen Synthase Kinase 3 beta (GSK3b) (AA 1-160) antibody

Antigen: Glycogen Synthase Kinase 3 beta (GSK3b)

Synonyms: GSK3B, GPR156, GSK3, GSK-3, C86142, GSK-3beta, 7330414F15Rik, 8430431H08Rik, GSK3beta, gsk3b, fk80d11, GSK-3[b], wu:fb68h05, wu:fk80d11, gsk3, gsk-3, xgsk-3, gsk3beta, MGC131076, gsk3-beta, ATSK42, F12K22.12, F12K22_12, shaggy-like kinase 42, SK42

Binding Site: AA 1-160

Clonality: Monoclonal (7)

Host: Mouse

Reactivity: Rat (Rattus)

Conjugate: Un-conjugated

Application: Western Blotting (WB), BioImaging (BI), Immunoprecipitation (IP), Immunohistochemistry (IHC)

Catalog no.: ABIN967820

Additional Information

Alternative name: GSK-3beta

Immunogen: Rat GSK-3beta

Cross-Reactivity: Mouse (Murine), Chicken, Dog (Canine), Human

Format: Liquid

Isotype: IgG1

Clone: 7

Description: Glycogen Synthase Kinase-3beta (GSK-3beta) is a serine/threonine kinase that affects glycogen metabolism by phosphorylating and, thus, down-regulating the activity of muscle glycogen synthase. GSK-3beta is identical to the Tau Protein Kinase 1 (TPK 1) which plays a role in the formation of the histopathological lesions of Alzheimer's disease (AD). In AD, many neurons contain intracytoplasmic neurofibrillary tangles (NFT) that are composed of ubiquitin and highly phosphorylated tau proteins, members of a family of microtubule-assocaited phosphoproteins. Phosphorylation of the tau proteins by GSK-3beta converts them into paired helical filaments (PHF) which are found in NFT and degenerative neurites of senile plaques. This ability of GSK-3beta to convert tau proteins to PHF has been demonstrated in vitro using primary cultures of embryonic rat hippocampal neurons.

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.

Molecular Weight: 46 kDa

Comments:

Related Products: ABIN967389

Synonyms: GSK3B, GPR156, GSK3, GSK-3, C86142, GSK-3beta, 7330414F15Rik, 8430431H08Rik, GSK3beta, gsk3b, fk80d11, GSK-3[b], wu:fb68h05, wu:fk80d11, gsk3, gsk-3, xgsk-3, gsk3beta, MGC131076, gsk3-beta, ATSK42, F12K22.12, F12K22_12, shaggy-like kinase 42, SK42

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Concentration: 250 µg/ml

Purity: Purified

Purification: Purified from tissue culture supernatant or ascites by affinity chromatography.

Buffer: Aqueous buffered solution containing BSA, glycerol.

Preservative: 0.09% Sodium azide.

Storage: Store undiluted at -20° C.

Restrictions: For Research Use only

Images

Western blot analysis of GSK-3beta on a RSV-3T3 lysate. Lane 1: 1:2500, lane 2: 1:5000, lane 3: 1:10000 dilution of the anti-GSK-3beta antibody.

Immunofluorescent staining of HeLa (ATCC CCL-2) cells. Cells were seeded in a 96 well imaging plate at ~10 000 cells per well. After overnight incubation, cells were stained using the alcohol perm protocol and the anti-GSK-3beta antibody. The second step reagent was Alexa Fluor® 555 goat anti-mouse IgG (Invitrogen). The image was taken on a BD Pathway™ 855 Bioimager using a 20x objective. This antibody also stained A549 (ATCC CCL-185) and U-2 OS (ATCC HTB-96) and cells and worked with both the Triton™ X-100 and alcohol perm protocols.

Publications

Product: Takashima, Noguchi, Sato et al.: "Tau protein kinase I is essential for amyloid beta-protein-induced neurotoxicity." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, Issue 16, pp. 7789-93, 1993 (PubMed).

Lucas, Hernández, Gómez-Ramos et al.: "Decreased nuclear beta-catenin, tau hyperphosphorylation and neurodegeneration in GSK-3beta conditional transgenic mice." in: The EMBO journal, Vol. 20, Issue 1-2, pp. 27-39, 2001 (PubMed).

Morisco, Seta, Hardt et al.: "Glycogen synthase kinase 3beta regulates GATA4 in cardiac myocytes." in: The Journal of biological chemistry, Vol. 276, Issue 30, pp. 28586-97, 2001 (PubMed).

Bijur, Jope: "Proapoptotic stimuli induce nuclear accumulation of glycogen synthase kinase-3 beta." in: The Journal of biological chemistry, Vol. 276, Issue 40, pp. 37436-42, 2001 (PubMed).

Etienne-Manneville, Hall: "Cdc42 regulates GSK-3beta and adenomatous polyposis coli to control cell polarity." in: Nature, Vol. 421, Issue 6924, pp. 753-6, 2003 (PubMed).