p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23)

Immunohistochemistry: p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) [NBP3-32783] - Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-p70 S6 Kinase/S6K antibody (NBP3-32783) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32783) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) [NBP3-32783] - Immunocytochemistry analysis of MCF7 cells treated with or without 20% FBS overnight then add 100nM Calyculin A for 30 minutes labeling p70 S6 Kinase/S6K with Rabbit anti-p70 S6 Kinase/S6K antibody (NBP3-32783) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-p70 S6 Kinase/S6K antibody (NBP3-32783) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Western Blot: p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) [NBP3-32783] - Western blot analysis of p70 S6 Kinase/S6K on different lysates with Rabbit anti-p70 S6 Kinase/S6K antibody (NBP3-32783) at 1/2,000 dilution. Lane 1: MCF7 cell lysate Lane 2: MCF7 treated with 20% FBS overnight then add 100nM Calyculin A for 30 minutes cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 treated with 20% FBS overnight then add 100nM Calyculin A for 30 minutes cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 59 kDa Observed band size: 70/85 kDa Exposure time: Lane 1-4 (left): 1 minute 2 seconds; Lane 1-4 (right): 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF, IHC
• Clone: PSH02-23
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) Summary

• Description: Novus Biologicals Rabbit p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23) (NBP3-32783) is a recombinant monoclonal antibody validated for use in IHC, WB and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Synthetic peptide within human p70 S6 Kinase/S6K aa 371-420 / 525. (Uniprot: P23443)
• Modification: p Thr389
• Localization: Synapse, synaptosome, Mitochondrion outer membrane, Mitochondrion.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: RPS6KB1
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:1000
  • Western Blot 1:1000-1:2000
• Theoretical MW: 59 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH7.4), 0.1% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for p70 S6 Kinase/S6K [p Thr389] Antibody (PSH02-23)

• EC 2.7.11
• EC 2.7.11.1
• p70 ribosomal S6 kinase alpha
• p70 S6 kinase alpha
• p70 S6 Kinase
• p70 S6 kinase, alpha 1,70 kDa ribosomal protein S6 kinase 1
• p70 S6 kinase, alpha 2
• p70 S6KA
• p70 S6K-alpha
• p70(S6K)-alpha
• p70-alpha
• p70-S6K
• P70S6K1
• PS6K
• ribosomal protein S6 kinase beta-1
• Ribosomal protein S6 kinase I
• ribosomal protein S6 kinase, 70kD, polypeptide 1
• ribosomal protein S6 kinase, 70kDa, polypeptide 1
• RPS6KB1
• S6K
• S6K1
• S6K1p70-S6K 1
• S6K-beta-1
• serine/threonine kinase 14 alpha
• Serine/threonine-protein kinase 14A
• STK14A

Background

S6K encodes a member of the RSK (ribosomal S6 kinase) family of serine/threonine kinases. This kinase contains 2 non-identical kinase catalytic domains and phosphorylates several residues of the S6 ribosomal protein. The kinase activity of this protein leads to an increase in protein synthesis and cell proliferation. Amplification of the region of DNA encoding this gene and overexpression of this kinase are seen in some breast cancer cell lines. Alternate translational start sites have been described and alternate transcriptional splice variants have been observed but have not been thoroughly characterized.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: RPS6KB1