Revision 8
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, W-S, IP, IHC-Bond, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M Mk

SENSITIVITY:

Endogenous

MW (kDa):

175

Source/Isotype:

Rabbit IgG

UniProt ID:

#P00533

Entrez-Gene Id:

1956

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:100
IHC Leica Bond 1:50
Immunohistochemistry (Paraffin) 1:50
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200
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 #26038.

Specificity / Sensitivity

EGF Receptor (D38B1) XP® Rabbit mAb detects endogenous levels of total EGF receptor protein. The antibody does not cross-react with other proteins of the ErbB family. Species cross-reactivity for IHC-P, IHC-BOND, and IF-IC is human only.

Species Reactivity:

Human, Mouse, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a fusion protein containing the cytoplasmic domain of human EGF receptor.

Background

The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, and lysosomal degradation (1,2). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme, and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for the adaptor protein c-Cbl, leading to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated EGFR residues (Tyr1148 and Tyr1173) provide a docking site for the Shc scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).

  1. Hackel, P.O. et al. (1999) Curr Opin Cell Biol 11, 184-9.
  2. Zwick, E. et al. (1999) Trends Pharmacol Sci 20, 408-12.
  3. Cooper, J.A. and Howell, B. (1993) Cell 73, 1051-4.
  4. Hubbard, S.R. et al. (1994) Nature 372, 746-54.
  5. Biscardi, J.S. et al. (1999) J Biol Chem 274, 8335-43.
  6. Emlet, D.R. et al. (1997) J Biol Chem 272, 4079-86.
  7. Levkowitz, G. et al. (1999) Mol Cell 4, 1029-40.
  8. Ettenberg, S.A. et al. (1999) Oncogene 18, 1855-66.
  9. Rojas, M. et al. (1996) J Biol Chem 271, 27456-61.
  10. Feinmesser, R.L. et al. (1999) J Biol Chem 274, 16168-73.

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 W-S: Simple Western™ IP: Immunoprecipitation IHC-Bond: IHC Leica Bond 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.
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 8
#4267

EGF Receptor (D38B1) XP® Rabbit mAb

Western Blotting Image 1: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb #4267(上图)或 #8457 β-Actin (D6A8) Rabbit mAb(下图)对对照 HeLa 细胞(泳道 1)或 EGFR 敲除型 HeLa 细胞(泳道 2)的提取物进行蛋白质印迹分析。EGFR 敲除型 HeLa 细胞中没有信号,这证实了抗体对 EGFR 的特异性。
Western Blotting Image 2: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb 对 A-431、BxPC3 和 HeLa 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 1: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb #4267 对 A-431细胞的裂解物 (0.1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的单一靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 66-440 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
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Immunohistochemistry Image 1: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 EGF Receptor (D38B1) Rabbit mAb 对石蜡包埋的人肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 1: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb 对石蜡包埋的人肝细胞癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb 对石蜡包埋的人肺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 3: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb 对石蜡包埋的人胎盘细胞进行免疫组织化学分析。
Immunohistochemistry Image 4: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb 对石蜡包埋的 MDA-MB-468(放大 EGFR,左)、HT-29(低 EGFR,中间)和 CAMA-1(EGFR 阴性,右)细胞进行免疫组织化学分析。
Immunofluorescence Image 1: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb(绿色)对未经(左)或经人表皮生长因子处理(右)的 A549 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Immunofluorescence Image 2: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb(绿色)对未经(左)或经人表皮生长因子处理(右)的 A549 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: EGF Receptor (D38B1) XP® Rabbit mAb Expand Image
使用 EGF Receptor (D38B1) XP® Rabbit mAb #4267(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对 Jurkat 细胞(蓝色)和 A431 细胞(绿色)进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。