Cell Signaling Technology

Product Pathways - Protein Translation

p70 S6 Kinase Substrates Antibody Sampler Kit #2903

eEF2 kinase   eef2k   eIF-4B   eif4   eIf4B   Eukaryotic elongation factor 2 kinase   eukaryotic translation initiation factor 4B   p70   p70-S6K   p70s6   ribosomal protein   S6   S6K   s6kinase  

REACTIVITY

No. Size Price
2903T 1 Kit ( 6 x 20 µl ) ¥5,301.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
p70 S6 Kinase (49D7) Rabbit mAb #2708 20 µl W,IHC-P, H,M,R,Mk, 70, 85 Rabbit
Phospho-eIF4B (Ser422) Antibody #3591 20 µl W, H,M,R,Mk, 80 Rabbit
Phospho-eEF2k (Ser366) Antibody #3691 20 µl W,IP, H,R,Mk, 105 Rabbit
Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb #4858 20 µl W,IHC-P,IHC-F,IF-IC,F, H,M,R,Mk,Mi,Sc, C,Pg, 32 Rabbit IgG
Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb #5364 20 µl W,IHC-P,IF-IC,F, H,M,R,Mk, C,Pg, 32 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl W, Goat
Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb #9234 20 µl W, H,M,R,Mk, C, 70, 85 Rabbit IgG

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.

p70 S6 Kinase Substrates Antibody Sampler Kit 里面的抗体都能够检测内源性的目标蛋白水平。 p70 S6 Kinase (49D7) Antibody #2708也识别 p85 S6 Kinase。Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb 兔单抗#9234 也识别Thr412位点磷酸化的 p85 S6 Kinase,还可能包括Thr401位点磷酸化的 S6KII。Kit里面的其他抗体不与其他蛋白交叉反应。

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.

抗体是由合成的针对人源的 S6 Kinase 氨基末端序列的肽段,及针对p70 S6 Kinase Thr389位点周围序列,核糖体蛋白Ser235和236位点 ,核糖体蛋白Ser240和244位点,eIF4B Ser422位点,eEF2k Ser366位点的磷酸化肽段免疫动物而生产的。多克隆抗体由protein A和肽亲和层析技术纯化。

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 four Western blot experiments.

p70 S6 Kinase Substrates Antibody Sampler Kit为您提供一个经济快速的评定 p70 S6激酶若干底物的方法。Kit包含足够进行四次Western blot 实验所需的一抗和二抗。

Western Blotting

Western Blotting

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.

Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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.

IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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).

IHC-P (paraffin)

IHC-P (paraffin)

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

Western Blotting

Western Blotting

Western blot analysis of extracts from PC12, NIH/3T3, and SK-N-MC cells, using p70 S6 Kinase (49D7) Rabbit mAb.

Western Blotting

Western Blotting

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).

IF-IC

IF-IC

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).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse spleen using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

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.

IHC-P (paraffin)

IHC-P (paraffin)

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.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

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).

Flow Cytometry

Flow Cytometry

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).

Western Blotting

Western Blotting

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).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb.

IHC-F (frozen)

IHC-F (frozen)

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

Western Blotting

Western Blotting

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 #9234 (upper), or p70 S6 Kinase (49D7) rabbit mAb #2708 (lower).

Western blot 方法检测细胞提取物:293,NIH/3T3和PC12细胞,血清饥饿处理,或者20%血清处理;使用的抗体是Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb 兔单抗#9234 (上图), or p70 S6 Kinase (49D7) rabbit mAb兔单抗 #2708 (下图)。

Western Blotting

Western Blotting

Western blot analysis of extracts from PC12 and NIH/3T3 cells, treated with λ phosphatase, FBS or PDGF as indicated, using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb #4858 (upper) or S6 Ribosomal Protein (5G10) Rabbit mAb #2217 (lower).

Western blot 方法检测细胞提取物:NIH/3T3和PC12细胞,λ磷酸酶, FBS 或 PDGF处理;使用的抗体是Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb 兔单抗#4858(上图), or S6 Ribosomal Protein (5G10) Rabbit mAb兔单抗 #2217 (下图)。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells using Phospho-eIF4B (Ser422) Antibody #3591 (upper) or eIF4B Antibody #3592 (lower). 48 hours following siRNA transfection, cells were treated with Rapamycin (50 nM) and U0126 (10 µM) as indicated.

Western blot 方法检测Hela细胞提取物,siRNA处理48小时后分别用Rapamycin (50 nM) 和U0126 (10 µM) 处理;使用的抗体是 Phospho-eIF4B (Ser422) Antibody #3591 (上图) 和eIF4B Antibody #3592 (下图)。

Western Blotting

Western Blotting

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 #3691 (upper) or eEF2k Antibody #3692 (lower).

Western blot 方法检测饥饿18小时的SW-13细胞提取物,用小牛肠碱性磷酸酶(CIP) 或20% 胎牛血清处理30分钟,及饥饿18小时的Hela细胞提取物,用200 nM TPA处理30分钟;使用的抗体是 Phospho-eEF2k (Ser366) Antibody #3691 (上图) 和eEF2k Antibody #3692 (下图)。

Western Blotting

Western Blotting

Western blot analysis of extracts from PC12, NIH/3T3, and SK-N-MC cells, using p70 S6 Kinase (49D7) Rabbit mAb #2708.

Western blot 方法检测细胞提取物:PC12, NIH/3T3 和 SK-N-MC细胞,使用的抗体是p70 S6 Kinase (49D7) Rabbit mAb兔单抗 #2708。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® p70/85 S6 Kinase siRNA I (+) 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.

IHC-P (paraffin)

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)

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)

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)

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)

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.

IF-IC

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).

Western Blotting

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).

Flow Cytometry

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.

Western Blotting

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 #5364 (upper) or S6 Ribosomal Protein (5G10) Rabbit mAb #2217 (lower).

Western blot 方法检测细胞提取物:MCF7 和 NIH/3T3细胞,100 nM insulin处理十分钟或者20% FBS 处理30分钟;使用的抗体是Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb 兔单抗#5364 (上图) or S6 Ribosomal Protein (5G10) Rabbit mAb 兔单抗#2217 (下图)。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse colon using Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

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).

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.

p70 S6激酶是有丝分裂原活化的丝氨酸/苏氨酸蛋白激酶,是细胞生长和G1期细胞周期正常进行必需的(1,2)。P70 S6激酶可以磷酸化核糖体40S亚基的S6蛋白并参与5‘ oligopyrimidine tract mRNAs的翻译控制(1)。核糖体蛋白S6的重要磷酸化位点存在于羧基末端的一小段含有Ser235, Ser236, Ser240, Ser244的区域(3,4)。 p70 S6激酶可以体内磷酸化修饰eIF4B的雷帕霉素敏感位点Ser422,体外翻译试验表明eIF4B Ser422位点突变为丙氨酸导致其活性显著降低(5)。 p70 S6 激酶和p90RSK磷酸化修饰eEF2K导致 eEF2K失活(6), 加速eEF2的去磷酸化最终促进翻译。

  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.

Application References

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

U.S. Patent No. 5,675,063.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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