Product Pathways - Translational Control
Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb #5364
PhosphoSitePlus® protein, site, and accession data: S6
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IHC-P IF-IC F | H M R Mk (C) (Pg) | Endogenous | 32 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
C=Chicken
Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb detects endogenous levels of ribosomal protein S6 only when phosphorylated at Ser240 and Ser244.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser240 and Ser244 of human ribosomal protein S6.
Western Blotting
Western blot analysis of extracts from MCF7 and NIH/3T3 cells, treated with 100 nM insulin (10 min) or 20% FBS (30 min) as indicated, using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb (upper) or S6 Ribosomal Protein (5G10) Rabbit mAb #2217 (lower).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded mouse colon using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cell pellets -/+ LY294002) using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded LNCaP cell pellets, control (left) or rapamycin-treated (right), using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded Rh30 xenograft, control (left) or rapamycin-treated (right), using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.
Flow Cytometry
Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002, wortmannin and U0126 (blue), using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.
IF-IC
Confocal immunofluorescent analysis of HeLa cells, insulin-treated (left) and LY294002-treated (#9901, right), using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 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).
- Dufner, A. and Thomas, G. (1999) Exp. Cell Res. 253, 100-109.
- Peterson, R.T. and Schreiber, S.L. (1998) Curr. Biol. 8, R248-R250.
- Jefferies, H.B. et al. (1997) EMBO J. 16, 3693-3704.
- Ferrari, S. et al. (1991) J. Biol. Chem. 266, 22770-22775.
- Flotow, H. and Thomas, G. (1992) J. Biol. Chem. 267, 3074-3078.
Application References
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Companion Products
- 8112 SignalStain® Antibody Diluent
- 8114 SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
- 8101 SignalSlide® Phospho-Akt (Ser473) IHC Controls
- 2217 S6 Ribosomal Protein (5G10) Rabbit mAb
- 4858 Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb
- 2708 p70 S6 Kinase (49D7) Rabbit mAb
- 9203 p70 S6 Kinase Control Cell Extracts
- 9204 Phospho-p70 S6 Kinase (Thr421/Ser424) Antibody
- 9234 Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb
- 9430 PhosphoPlus® p70 S6 Kinase (Thr389, Thr421/Ser424) Antibody Kit
- 9918 Translational Control Antibody Sampler Kit
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
