Detection of p70 S6 Kinase in HeLa Human Cell Line by Flow Cytometry.
HeLa human cervical epithelial carcinoma cell line was stained with Human/Mouse/Rat p70 S6 Kinase Monoclonal Antibody (Catalog # MAB8962, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.
Detection of Human/Mouse/Rat p70 S6 Kinase by Western Blot.
Western blot shows lysates of MCF-7 human breast cancer cell line, PC-12 rat adrenal pheochromocytoma cell line, and TS1 mouse helper T cell line. PVDF membrane was probed with 0.25 µg/mL Human/Mouse/Rat p70 S6 Kinase Monoclonal Antibody (Catalog # MAB8962) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, Recombinant Human Active p70 S6 Kinase (Catalog # 896-KS) was included. A specific band for p70 S6 Kinase was detected at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: p70 S6 Kinase
p70 S6 Kinase (p70S6K) is responsible for the phosphorylation of 40S ribosomal protein S6 and is ubiquitously expressed in human adult tissues (1). p70S6K is activated by serum stimulation and this activation is inhibited by wortmannin and rapamycin. p70S6K activity undergoes changes in the cell cycle and increases 20‑fold in G1 cells released from G0 (2). p70S6K activation requires sequential phosphorylations at proline-directed residues in the putative autoinhibitory pseudosubstrate domain, as well asT389, a site phosphorylated by phosphoinositide-dependent kinase 1 (PDK1).
Ferrari, S. et al. (1994) Crit. Rev. Biochem. Mol. Biol. 29:385.
Edelmann, H.M. et al. (1996) J. Biol. Chem. 271:963.
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