Product Name: p70S6K-pT252
Product Number: AB-PK744
Size: 25 µg      Price:89.00
      $US
Target Full Name: Ribosomal protein S6 kinase beta-1; Ribosomal protein S6 kinase 1

Target Alias: RPS6KB1; KS6B1; Ps6K; p70(S6K)-alpha; Ribosomal protein S6 kinase I; RPS6KB1; S6K; S6K1; STK14A

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK744

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: T252

Protein UniProt: P23443

Protein SigNET: P23443

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human p70S6K (S6Ka, RPS6KB1) sequence peptide Cat. No.: PE-04AKT99

Antibody Immunogen Sequence: VTH(pT)FCG(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues V249 to G255; In protein kinase catalytic domain activation T-loop between subdomains VII and VIII.

Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.

Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.

Antibody Concentration: 1 mg/ml

Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol

Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.

Product Use: Western blotting | Antibody microarray

Antibody Dilution Recommended: 2 µg/ml for immunoblotting

Antibody Potency: Weak immunoreactivity of a target-sized protein by Western blotting in MCF7 cells. Medium immunoreactivity with immunogen peptide on dot blots.

Antibody Species Reactivity: Human

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 63-70 kDa.

Antibody Specificity: Very high

Antibody Cross Reactivity: Almost no significant cross-reactivities detected in HepG2 and T98G cells, and in HeLa cells, phenylarsine oxide (PAO) decreases detection of 260 + 75 kDa proteins.

Related Product 1: p70S6K-pT252 blocking peptide

Related Product 2: p70S6K-pT412 phosphosite-specific antibody (Cat. No.: AB-PK745)

Related Product 3: p70S6K-pT444+pS447 phosphosite-specific antibody (Cat. No.: AB-PK746)

Related Product 4: RPS6 (230-238) KinSub - 40S ribosomal protein S6 (K230-R238, human) peptide; p70S6K protein kinase substrate peptide

Related Product 5: S6KASubtide - p70S6K (RPS6KB1) protein kinase substrate peptide

Related Product 6: GSK3b (3-12) KinSub - Glycogen synthase kinase 3-beta (GSK3b) N-terminus (G3-E12, human) peptide; Akt protein kinase substrate peptide

Scientific Background: p70S6K (RPS6KB1) is a protein-serine/threonine kinase that is a member of the AGC group of protein kinases in the RSK family, and p70 subfamily. This kinase is highly expressed and widely distributed in most tested human tissues except in the brain and spinal cord where it is more moderately expressed. It functions downstream of mTOR (FRAP1) signalling in response to growth factors to promote cell proliferation, growth, and progression through the cell cycle. p70S6K is activated by phosphorylation at T252 (by PDK1), S394 (by mTOR), T412 (by mTOR), S427, S434 (by ERK1/2), S441, T444 (by ERK1/2) and S447 (by ERK1/2). p70S6K is responsible for the phosphorylation of 40S ribosomal protein S6. It is activated by serum stimulation and this activation is inhibited by wortmannin and rapamycin. p70S6K phosphotransferase activity changes during the cell cycle, and increases 20-fold in G1 cells released from G0. It has been linked with the development of diet-induced obesity, colon adenocarcinoma and breast cancer.
Figure 1. Epitope mapping of p70S6K-pT252 antibody with similar phosphopeptides on dot blots.


Figure 2. Identification of phosphosites related to p70S6K-pT252.