Product Name:
RPS6-pS235+pS236+pS240
Product Number:
ab-pn559
Target Full Name: 40S Ribosomal S6
Target Alias: Phosphoprotein NP33; Ribosomal protein S6; RS6; S6 Ribosomal Protein
Product Type Specific: RPS6 phosphosite-specific antibody
Antibody Code: PN559
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S235+S236+S240
Protein UniProt: P62753
Protein SigNET: RPS6
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human RPS6 sequence peptide Cat. No.: PE-04AQR95
Antibody Immunogen Sequence: RRL(pS)(pS)LRA(pS)TS(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues R232 to S242; in the C-terminus of the ribosomal protein. S235+S236+S240 phosphorylation is believed to increase the translation of mRNA important for protein synthesis by facilitating the assembly of the preinitiation complex. These are the major sites of in vivo phosphorylation of S6 based on ≥1030, ≥1194 and ≥916 high throughput mass spectrometry reports, respectively, recorded in PhosphoSitePlus. RPS6 is known to be phosphorylated in vitro at S235 by Akt1 (PKBa), Akt2 (PKBb), AurB (STK12), DAPK1, p70S6K (RPS6KB1), PASK, PKACa (PRKACA), PKCd (PRKCD), RSK1 (RPS6KA1), and RSK2 (RPS6KA3), at S236 by Akt1 (PKBa), Akt2 (PKBb), AurB (STK12), DAPK1, p70S6K (RPS6KB1), PASK, PKACa (PRKACA), PKCa (PRKCA), PKCd (PRKCD), RSK1 (RPS6KA1), and RSK2 (RPS6KA3), and at S240 by p70S6K (RPS6KB1), and PKACa (PRKACA).
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.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.1 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: Medium immunoreactivity of a target-sized protein by Western blotting in MCF7 + insulin (homogenized in SDS-PAGE buffer).
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Dog | Rat | Mouse | Platypus | Chicken | Frog | Zebra fish.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 28-34 kDa.
Antibody Specificity: Very high
Antibody Cross Reactivity: No significant cross-reactive proteins detected in phenylarsine oxide (PAO)+vanadate-treated HeLa cells, EGF-treated A431 cells and insulin-treated MCF7 cells, when these cells were homogenized in SDS-PAGE sample buffer.
Related Product 1: RPS6-pS235+pS236+pS240 blocking peptide
Related Product 2: RPS6 (230-238) KinSub - 40S ribosomal protein S6 (K230-R238, human) peptide; p70S6K protein kinase substrate peptide
Related Product 3: RPS6 (230-239) KinSub kinase substrate peptide (Cat. No.: PE-01ADP95)
Related Product 4: RPS6 S235+S236 phosphorylation antibody (Cat. No.: AB-PN685)
Related Product 5: RPS6 S240+S244 phosphorylation antibody (Cat. No.: AB-PN1025)
Scientific Background: RPS6 (Ribosomal protein S6; eS6) is a key 32–34 kDa component of the 40S ribosomal subunit in eukaryotes that plays a vital role in protein synthesis and translation regulation. It controls cell growth and proliferation through the selective translation of particular classes of mRNA that are important in protein synthesis. RPS6 is located in the small head region of the 40S subunit, at the interface with the 60S subunit. While it is a structural component of the ribosome, its phosphorylation is not required for general translation, but rather specifically affects the translation efficiency of specific mRNA classes Growth factor stimulation of cultured cells leads to stong phosphorylation of RPS6 at multiple sites located in its C-terminus (specifically serine residues 235, 236, 240, 244, and 247 in mammals), which enhances it translational activity. Phosphorylation is generally triggered by mitogenic stimuli, nutrients, and growth factors via the mTOR pathway. This phosphorylation may be mediated by several protein kinases including those in the p70 S6 kinase and RSK families. RPS6 acts as a regulator of cell size, proliferation, and glucose homeostasis. This description may include information annotated by UniProt and/or Google AI.