Scientific Data

Western Blot

Detection of Human/Mouse/Rat GSK‑3 beta antibody by Western Blot.

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Detection of Human/Mouse/Rat GSK‑3 beta by Western Blot. Western blot shows lysates of HT-29 human colon adenocarcinoma cell line and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 1 µg/mL Human/Mouse/Rat GSK-3 beta Monoclonal Antibody (Catalog # MAB2506) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). For additional reference, recombinant human GSK-3a and Recombinant Human Active GSK-3 beta (Catalog # 2506-KS) (20 ng/lane) were included. A specific band for GSK-3 beta was detected at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

GSK-3 beta antibody in HT-29 Human Cell Line by Immunocytochemistry (ICC).

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GSK‑3 beta in HT‑29 Human Cell Line. GSK-3 beta was detected in immersion fixed HT-29 human colon adenocarcinoma cell line using Rat Anti-Human/Mouse/Rat GSK-3 beta Monoclonal Antibody (Catalog # MAB2506) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunocytochemistry

GSK-3 beta antibody in NIH-3T3 Mouse Cell Line by Immunocytochemistry (ICC).

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GSK‑3 beta in NIH‑3T3 Mouse Cell Line. GSK-3 beta was detected in immersion fixed NIH-3T3 mouse embryonic fibroblast cell line using Rat Anti-Human/Mouse/Rat GSK-3 beta Monoclonal Antibody (Catalog # MAB2506) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western Blot

Detection of Mouse GSK-3 beta by Western Blot

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Detection of Mouse GSK-3 beta by Western Blot BSSM activates AKT/GSK3 beta pathway, promotes tau hyperphosphorylation and PHF accumulation and increases the expression of active asparagine endopeptidase (AEP) and tau cleavage. (A,B) Western blotting showed higher expressions of p-AKT and the downstream p-GSK in hippocampus in BSSM treated group. (C,D) Phosphorylation of tau at S396 was higher in BSSM group than in sham and USSM group. PHF accumulation also showed significant differences between sham and the two shear stress modifier groups. (E–G) Expression of active AEP and tau N368 truncation were higher in BSSM group. (H,I) Immunofluorescent staining further reconfirmed the finding that BSSM group had more tau N368 fragments in hippocampus than the other two groups. L, left hippocampus; R, right hippocampus. *p < 0.05, **p < 0.01. Data were presented as Mean ± SD. N = 4 in each group. Scale bar, 100 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30337867), licensed under a CC-BY license. Not internally tested by R&D Systems.

Human/Mouse/Rat GSK-3 beta Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human, mouse, and rat GSK-3 beta in ELISAs and Western blots. Does not cross-react with recombinant human GSK-3 alpha.

Source

Monoclonal Rat IgG_2B Clone # 272536

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human GSK-3 beta
Met1-Thr420
Accession # P49841

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

3-25 µg/mL

See below

Intracellular Staining by Flow Cytometry

2.5 µg/10⁶ cells

HeLa human cervical epithelial carcinoma cell line fixed with paraformaldehyde and permeabilized with methanol.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

h/m/rGSK-3b MAb (Cl (25 ug)