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2903
p70 S6 Kinase Substrates Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

p70 S6 Kinase Substrates Antibody Sampler Kit #2903

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p70 S6 Kinase Substrates Antibody Sampler Kit: Image 1
Western blot analysis of extracts from PC12, NIH/3T3, and SK-N-MC cells, using p70 S6 Kinase (49D7) Rabbit mAb.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 2
Western blot analysis of extracts from HeLa cells using Phospho-eIF4B (Ser422) Antibody (upper) or eIF4B Antibody #3592 (lower). 48 hours following siRNA transfection, cells were treated with Rapamycin (50 nM) and U0126 (10 µM) as indicated.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 3
Western blot analysis of extracts from SW-13 cells (starved for 18 hours) treated with calf intestinal alkaline phosphatase (CIP) or 20% fetal bovine serum for 30 minutes, and extracts from HeLa cells (starved for 18 hours) treated with 200 nM TPA for 30 minutes, using Phospho-eEF2k (Ser366) Antibody (upper) or eEF2k Antibody #3692 (lower).
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 4
Western blot analysis of extracts from PC12 and NIH/3T3 cells, treated with λ phosphatase, 20% FBS (20 min) or 100 ng/ml PDGF (20 min) as indicated, using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (upper) or S6 Ribosomal Protein (5G10) Rabbit mAb #2217 (lower).
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 5
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).
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 6
Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, wortmannin #9951 and U0126 #9903 (50 μM, 1 μM and 10 μM, 2hr; blue) using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 7
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 8
Western blot analysis of extracts from serum starved or serum treated (20%) 293, NIH/3T3, and PC12 cells, using Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb (upper), or p70 S6 Kinase (49D7) rabbit mAb #2708 (lower).
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 9
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® p70/85 S6 Kinase siRNA I #6566 (+) or SignalSilence® p70/85 S6 Kinase siRNA II #6572 (+), using p70 S6 Kinase (49D7) Rabbit mAb #2708 and α-Tubulin (11H10) Rabbit mAb #2125. The p70 S6 Kinase (49D7) Rabbit mAb confirms silencing of p70/85 S6 kinase expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p70/85 S6 kinase siRNA.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 10
Immunohistochemical analysis using Phospho-S6 Ribosomal Protein (S235/236) (D57.2.2E) XP® Rabbit mAb on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right).
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 11
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).
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 12
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb in the presence of control peptide (left) or Phospho-S6 Ribosomal Protein (Ser235/236) Blocking Peptide #1220 (right).
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 13
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 14
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 15
Immunohistochemical analysis of paraffin-embedded mouse colon using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 16
Immunohistochemical analysis of paraffin-embedded LNCaP cells, untreated (left) or rapamycin-treated (right), using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 17
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.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 18
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 19
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.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded mouse spleen using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 21
Immunohistochemical analysis of paraffin-embedded Rh30 xenograft, control (left) or rapamycin-treated (right), using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 22
Immunohistochemical analysis of paraffin-embedded A549 xenograft, untreated (left) or λ phosphatase-treated (right), using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb.
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 23
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).
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 24
Confocal immunofluorescent analysis of HeLa cells, rapamycin-treated (left) or 20% serum-treated (right), using Phospho-S6 Ribosomal protein (Ser235/Ser236) (D57.2.2E) XP® Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
p70 S6 Kinase Substrates Antibody Sampler Kit: Image 25
Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, Wortmannin #9951, and U0126 #9903 (50 μM, 1 μM, and 10 μM, 2 hr; blue) using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
To Purchase # 2903
Cat. # Size Qty. Price
2903T
1 Kit  (6 x 20 microliters) $ 458

Product Description

The p70 S6 Kinase Substrates Antibody Sampler Kit provides a fast and economical means of evaluating several substrates of p70 S6 Kinase. The kit contains enough primary and secondary antibody to perform two Western blot experiments.

Specificity / Sensitivity

Each antibody in the p70 S6 Kinase Substrates Antibody Sampler Kit detects endogenous levels of its target protein. p70 S6 Kinase (49D7) Antibody #2708 also recognizes p85 S6 Kinase. Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb #9234 also detects p85 S6 Kinase when phosphorylated at Thr412 and possibly S6KII when phosphorylated at Thr401. The other antibodies in the kit do not cross react with other proteins.

Source / Purification

Antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the amino-terminus of human p70 S6 Kinase; and to synthetic phosphopeptides corresponding to residues surrounding Thr389 of human p70 S6 Kinase; Ser235 and 236 of human ribosomal protein; Ser240 and 244 of human ribosomal protein; Ser422 of human eIF4B; and Ser366 of human eEF2k. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

p70 S6 kinase is a mitogen activated Ser/Thr protein kinase that is required for cell growth and G1 cell cycle progression (1,2). p70 S6 kinase phosphorylates the S6 protein of the 40S ribosomal subunit and is involved in translational control of 5' oligopyrimidine tract mRNAs (1). Important S6 ribosomal protein phosphorylation sites include several residues (Ser235, Ser236, Ser240, Ser244) located wtihin a small, carboxy-terminal region of the S6 protein (3,4). p70 S6 kinase has been shown to phosphorylate eIF4B at the rapamycin-sensitive site Ser422 in vivo, and a Ser422Ala mutant of eIF4B shows diminished activity in an in vitro translation assay (5). Phosphorylation of eEF2K by p70 S6 kinase and p90RSK leads to inactivation of eEF2K (6), facilitating the dephosphorylation of eEF2 and thus promoting translation.
  1. Pullen, N. and Thomas, G. (1997) FEBS Lett 410, 78-82.
  2. Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9.
  3. Ferrari, S. et al. (1991) J Biol Chem 266, 22770-5.
  4. Flotow, H. and Thomas, G. (1992) J Biol Chem 267, 3074-8.
  5. Raught, B. et al. (2004) EMBO J 23, 1761-9.
  6. Wang, X. et al. (2001) EMBO J 20, 4370-9.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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