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Phospho-S6 Ribosomal Protein (Ser235/236) Antibody #2211

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PhosphoSitePlus^® protein, site, and accession data: S6

Applications	Reactivity	Sensitivity	MW (kDa)	Source
W IP IHC-P IHC-F IF-IC F	H M R Mk Sc (C) (X)	Endogenous	32	Rabbit

Applications Key: W=Western Blotting IP=Immunoprecipitation IHC-P=Immunohistochemistry (Paraffin) IHC-F=Immunohistochemistry (Frozen) IF-IC=Immunofluorescence (Immunocytochemistry) F=Flow Cytometry
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey C=Chicken X=Xenopus Sc=S. cerevisiae
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

2211:: Flow, IHC / Frozen, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

Specificity / Sensitivity

Phospho-S6 Ribosomal Protein (Ser235/236) Antibody detects endogenous levels of ribosomal protein S6 only when phosphorylated at serine 235 and 236. This antibody does not detect ribosomal protein S6 phosphorylated at other sites.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser235 and Ser236 of human ribosomal protein S6. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or treated with 20% FBS for the indiccated time, using , Phospho-S6 Ribosomal Protein (Ser235/236) Antibody (upper) or Phospho-S6 Ribosomal Protein (Ser240/244) Antibody #2215 (lower).

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human melanoma, showing cytoplasmic localization of phosphorylated S6 ribosomal protein, using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or calf-intestinal phosphatase (CIP) treated (right), using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma, using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded LNCaP cells, untreated (left) or rapamycin-treated (right), using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody.

IHC-P (paraffin)

Imunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody preincubated with control peptide (left) or Phospho-S6 Ribosomal Protein (Ser235/236) Blocking Peptide #1220 (right).

IHC-F (frozen)

Immunohistochemical analysis of frozen U87MG xenograft, using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody.

Flow Cytometry

Flow cytometric analysis of NIH/3T3 cells, U0126- and Rapamycin-treated (blue) or serum-treated (green), using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody compared to a nonspecific negative control antibody (red).

IF-IC

Immunofluorescent analysis of HeLa cells, untreated (A), 20% serum-treated (B) and serum-treated after rapamycin pretreatment (C), using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody.

IF-IC

Confocal immunofluorescent analysis of HeLa cells, serum-starved (left) or 20% serum-treated (right), using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Background

One way that growth factors and mitogens effectively promote sustained cell growth and proliferation is by upregulating mRNA translation (1,2). Growth factors and mitogens induce the activation of p70 S6 kinase and the subsequent phosphorylation of the S6 ribosomal protein. Phosphorylation of S6 ribosomal protein correlates with an increase in translation of mRNA transcripts that contain an oligopyrimidine tract in their 5' untranslated regions (2). These particular mRNA transcripts (5'TOP) encode proteins involved in cell cycle progression, as well as ribosomal proteins and elongation factors necessary for translation (2,3). Important S6 ribosomal protein phosphorylation sites include several residues (Ser235, Ser236, Ser240, and Ser244) located within a small, carboxy-terminal region of the S6 protein (4,5).

Application References

Tang, H. et al. (2001) Amino acid-induced translation of TOP mRNAs is fully dependent on phosphatidylinositol 3-kinase-mediated signaling, is partially inhibited by rapamycin, and is independent of S6K1 and rpS6 phosphorylation. Mol. Cell. Biol. 21, 8671-8683. Applications: Western Blot
Neshat, M. S. et al. (2001) Enhanced sensitivity of PTEN-deficient tumors to inhibition of FRAP/mTOR. Proc. Natl. Acad. Sci. USA 98, 10314-10319. Applications: Western Blot
Kenerson, H. L. et al. (2002) Activated mammalian target of rapamycin pathway in the pathogenesis of tuberous sclerosis complex renal tumors. Cancer Res. 62, 5645-5650. Applications: IHC-P (paraffin) Western Blotting
Castro, A. F. et al. (2003) Rheb binds tuberous sclerosis complex 2 (TSC2) and promotes S6 kinase activation in a rapamycin- and farnesylation-dependent manner. J. Biol. Chem. 278, 32493-32496. Applications: Western Blotting
Choe, G. et al. (2003) Analysis of the phosphatidylinositol 3'-kinase signaling pathway in glioblastoma patients in vivo. Cancer Res. 63, 2742-2746. Applications: IHC-P (paraffin)
Fonseca, B.D. et al. (2011) J Biol Chem 286, 27111-22. Applications: Western Blotting
Pitter, K.L. et al. (2011) PLoS One 6, e14545. Applications: IHC-P (paraffin)
De Raedt, T. et al. (2011) Cancer Cell 20, 400-13. Applications: Western Blotting

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