Detection of Human p70 S6 Kinase beta by Western Blot.
Western blot shows lysates of MCF‑7 human breast cancer cell line. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Human p70 S6 Kinase beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2987) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for p70 S6 Kinase beta at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
p70 S6 Kinase beta in MCF‑7 Human Cell Line.
p70 S6 Kinase beta was detected in immersion fixed MCF‑7 human breast cancer cell line using Goat Anti-Human p70 S6 Kinase beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2987) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
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 beta
P70S6K beta (p70 ribosomal S6 kinase beta-2; also RPS6KB2, S6K beta, S6K beta 2, S6K2 and STK14B) is a monomeric, 54 kDa member of the AGC family of Ser/Thr kinases. It is widely expressed, and found predominately in the nucleus. Although it was initially reported to phosphorylate the ribosomal S6 protein, p70S6K beta /S6K beta 2 is now known to also phosphorylate RafB (cell survival), hnRNP (cell proliferation), p300/CBP (transcription) and CDC42 (cytoskeleton stability). Human p70S6K beta /S6K beta 2 is 482 amino acids in length. It contains a protein kinase domain (aa 67-328), a C-terminal regulatory region (aa 329-399) that contains an NLS (aa 384-389), and a Pro-rich segment (aa 411-482) that likely interacts with SH3 domain-containing proteins. Phosphorylation/dephosphorylation events and sumoylation regulate p70S6K beta activity. In addition to the 54 kDa form, there is a 56-60 kDa isoform that possesses a 13 aa addition at the N-terminus, plus an isoform that shows a deletion of aa 267-270. Over aa 1-410, human p70S6K beta 2 shares 95% and 75% aa identity with mouse p70S6K beta 2and human p70S6K beta 1, respectively.
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