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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

Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002, wortmannin and U0126 (blue), using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb compared to a nonspecific negative control antibody (red).

p70 S6 Kinase Substrates Antibody Sampler Kit: Image 2

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.

p70 S6 Kinase Substrates Antibody Sampler Kit: Image 3

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 4

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 5

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 6

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using p70 S6 Kinase (49D7) Rabbit mAb.

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

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 9

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 10

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, showing cytoplasmic and nuclear localization, using p70 S6 Kinase (49D7) Rabbit mAb.

p70 S6 Kinase Substrates Antibody Sampler Kit: Image 11

Immunohistochemical analysis of frozen U-87MG xenograft using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb.

p70 S6 Kinase Substrates Antibody Sampler Kit: Image 12

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 13

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using p70 S6 Kinase (49D7) Rabbit mAb in the presence of control peptide (left) or p70 S6 Kinase Blocking Peptide #1205 (right).

p70 S6 Kinase Substrates Antibody Sampler Kit: Image 14

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 15

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 16

Immunohistochemical analysis of paraffin-embedded mIMCD-3 Tsc1 (-/-) cell pellet (left, positive) or mIMCD-3 Tsc1/S6k1 (-/-) cell pellet (right, negative) using p70 S6 Kinase (49D7) Rabbit mAb. (Cell lines courtesy of Dr. Mario Pende, Inserm, Paris, France.)

p70 S6 Kinase Substrates Antibody Sampler Kit: Image 17

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 18

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 19

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 20

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 21

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 22

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 23

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 24

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 25

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 26

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 27

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 28

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 29

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 30

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).

To Purchase # 2903T
Product # Size Price
2903T
1 Kit  (6 x 20 µl) $ 445

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb 4858 20 µl
  • WB
  • IHC
  • IF
  • F
H Mi Mk M R Sc 32 Rabbit IgG
Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb 5364 20 µl
  • WB
  • IHC
  • IF
  • F
H Mk M R 32 Rabbit IgG
Phospho-eIF4B (Ser422) Antibody 3591 20 µl
  • WB
H Mk M R 80 Rabbit 
Phospho-eEF2k (Ser366) Antibody 3691 20 µl
  • WB
  • IP
H Mk R 105 Rabbit 
Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb 9234 20 µl
  • WB
H Mk M R 70, 85 Rabbit IgG
p70 S6 Kinase (49D7) Rabbit mAb 2708 20 µl
  • WB
  • IHC
H Mk M R 70, 85 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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, 4370-4379.

Limited Uses

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