Revision 7
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M R Mk Dm

SENSITIVITY:

Endogenous

MW (kDa):

15 to 20

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q13541

Entrez-Gene Id:

1978

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:800 - 1:3200
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #39788.

Specificity / Sensitivity

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb detects endogenous levels of 4E-BP1 only when phosphorylated at Thr37 and/or Thr46. This antibody may cross-react with 4E-BP2 and 4E-BP3 when phosphorylated at equivalent sites. Non-specific staining has been observed in mitotic cells by immunofluorescence.

Species Reactivity:

Human, Mouse, Rat, Monkey, D. melanogaster

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr37 and Thr46 of mouse 4E-BP1.

Background

Translation repressor protein 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation by binding to the translation initiation factor eIF4E. Hyperphosphorylation of 4E-BP1 disrupts this interaction and results in activation of cap-dependent translation (1). Both the PI3 kinase/Akt pathway and FRAP/mTOR kinase regulate 4E-BP1 activity (2,3). Multiple 4E-BP1 residues are phosphorylated in vivo (4). While phosphorylation by FRAP/mTOR at Thr37 and Thr46 does not prevent the binding of 4E-BP1 to eIF4E, it is thought to prime 4E-BP1 for subsequent phosphorylation at Ser65 and Thr70 (5).

  1. Pause, A. et al. (1994) Nature 371, 762-7.
  2. Brunn, G.J. et al. (1997) Science 277, 99-101.
  3. Gingras, A.C. et al. (1998) Genes Dev 12, 502-13.
  4. Fadden, P. et al. (1997) J Biol Chem 272, 10240-7.
  5. Gingras, A.C. et al. (1999) Genes Dev 13, 1422-37.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 7
#2855

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb

Western Blotting Image 1: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb Expand Image
使用 4E-BP1 Antibody #9452(上)和 Phospho-4E-BP1 (Thr37/46) Antibody #2855(下)对来自 293T 细胞的提取物进行蛋白质印迹分析。细胞在无血清培养基中饥饿 24 小时并且给予氨基酸剥夺处理 1 小时。补充氨基酸 1 小时。随后,不处理 (-) 或用 100 nM 胰岛素处理 (+) 细胞 30 分钟。
Immunohistochemistry Image 1: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb Expand Image
使用 Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb 对石蜡包埋的人结肠癌进行免疫组织化学分析。
Immunohistochemistry Image 2: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb Expand Image
使用 Phospho-4E-BP1 (Thr37/46) (236B4) 兔单抗对石蜡包埋的人淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 3: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb Expand Image
使用 Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb 对 SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101(未经处理(左图)或经 LY294002 处理(右图)的石蜡包埋的 LNCaP 细胞)进行免疫组织化学分析。
Immunohistochemistry Image 4: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb Expand Image
在对照肽(左)或 Phospho-4E-BP1 (Thr37/46) Blocking Peptide #1052(右)的存在下,使用 Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb 对石蜡包埋的人结肠癌进行免疫组织化学分析。
Immunofluorescence Image 1: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb Expand Image
使用 Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb(绿色),对表达非靶向性 shRNA(底部)或靶向 4E-BP1/2 的 shRNA(顶部)的 293 细胞进行共聚焦免疫荧光分析。为证实磷酸特异性,细胞以 LY294002 #9901、U0126 #9903 和 Rapamycin #9904(50 μM;10 μm;10 nM;2小时)(左)组成的抑制剂混合物处理、胰岛素 (100 nM,30 分钟;中间)刺激、或胰岛素刺激后(右)用λ-磷酸酶处理。红色 = Propidium Iodide (PI)/RNase Staining Solution (#4087)。
Flow Cytometry Image 1: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb Expand Image
使用 Phospho-4E-BP1 (Thr36/46) (236B4) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对未经处理(绿色)或已经过 LY294002 #9901、Wortmannin #9951 和 U0126 #9903(50 μM、1 μM 和 10 μM,2 小时;蓝色)处理的 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。