Revision 5
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

SENSITIVITY:

Endogenous

MW (kDa):

185

Source/Isotype:

Rabbit IgG

UniProt ID:

#P21860

Entrez-Gene Id:

2065

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:50
IHC Leica Bond 1:50
Immunohistochemistry (Paraffin) 1:250
Immunofluorescence (Immunocytochemistry) 1:200
Flow Cytometry (Fixed/Permeabilized) 1:100

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 #60638.

Specificity / Sensitivity

HER3/ErbB3 (D22C5) XP® Rabbit mAb recognizes endogenous levels of total HER3/ErbB3 protein. This antibody does not cross-react with other HER family proteins.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein corresponding to the carboxy terminus of human ErbB3 protein.

Background

HER3/ErbB3 is a member of the ErbB receptor protein tyrosine kinase family, but it lacks tyrosine kinase activity. Tyrosine phosphorylation of ErbB3 depends on its association with other ErbB tyrosine kinases. Upon ligand binding, heterodimers form between ErbB3 and other ErbB proteins, and ErbB3 is phosphorylated on tyrosine residues by the activated ErbB kinase (1,2). There are at least 9 potential tyrosine phosphorylation sites in the carboxy-terminal tail of ErbB3. These sites serve as consensus binding sites for signal transducing proteins, including Src family members, Grb2, and the p85 subunit of PI3 kinase, which mediate ErbB downstream signaling (3). Both Tyr1222 and Tyr1289 of ErbB3 reside within a YXXM motif and participate in signaling to PI3K (4).

Investigators have found that ErbB3 is highly expressed in many cancer cells (5) and activation of the ErbB3/PI3K pathway is correlated with malignant phenotypes of adenocarcinomas (6). Research studies have demonstrated that in tumor development, ErbB3 may function as an oncogenic unit together with other ErbB members (e.g., ErbB2 requires ErbB3 to drive breast tumor cell proliferation) (7). Thus, investigators view inhibiting interaction between ErbB3 and ErbB tyrosine kinases as a novel strategy for anti-tumor therapy.

  1. Yarden, Y. and Sliwkowski, M.X. (2001) Nat Rev Mol Cell Biol 2, 127-37.
  2. Guy, P.M. et al. (1994) Proc Natl Acad Sci U S A 91, 8132-6.
  3. Songyang, Z. et al. (1993) Cell 72, 767-78.
  4. Kim, H.H. et al. (1994) J Biol Chem 269, 24747-55.
  5. Sithanandam, G. et al. (2003) Carcinogenesis 24, 1581-92.
  6. Kobayashi, M. et al. (2003) Oncogene 22, 1294-301.
  7. Holbro, T. et al. (2003) Proc Natl Acad Sci U S A 100, 8933-8.

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 nonfat dry milk, 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.
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 5
#12708

HER3/ErbB3 (D22C5) XP® Rabbit mAb

Western Blotting Image 1: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对 MCF7 (HER3+) 和 MDA-MB-231 (HER3-) 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 1: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
Simple Western™ analysis of lysates (1.0 mg/mL) from MCF-7 cells using HER3/ErbB3 (D22C5) XP® Rabbit mAb #12708. 虚拟泳道式图像(左图)显示一抗稀释比例为 1:50 和 1:250 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:50(蓝线)和 1:250(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 44-660 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
No image available
Immunohistochemistry Image 1: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
在 Leica BOND Rx 上使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb 对石蜡包埋的人导管乳腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 2: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
在 Leica BOND Rx 上使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb 对石蜡包埋的人卵巢透明细胞癌组织进行免疫组织化学分析。
Immunohistochemistry Image 3: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
在 Leica BOND Rx 上使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb 对石蜡包埋的人胃癌组织进行免疫组织化学分析。
Immunohistochemistry Image 1: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb 对石蜡包埋的非小细胞肺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb 对石蜡包埋的乳腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 3: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb 对石蜡包埋的卵巢浆液性腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 4: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb(顶板)对经人 erbB 家族成员 HER3、EGFR、HER2 和 HER4(根据具体情况,从左到右)转染的石蜡包埋的 293T 细胞团进行免疫组织化学分析。使用 EGF Receptor (D38B1) XP® Rabbit mAb #4267(左下图)、HER2/ErbB2 (D8F12) XP® Rabbit mAb #4290(中下图)和 HER4/ErbB4 (111B2) Rabbit mAb #4795(右下图)确认转染情况。
Immunohistochemistry Image 5: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb 对石蜡包埋的 MCF7(左图)或 MDA-MB-231(右图)细胞团进行免疫组织化学分析。
Immunofluorescence Image 1: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
使用 HER3/ErbB3 (D22C5) XP® Rabbit mAb(绿色)对 MCF7 细胞(HER3+,左图)和 MDA-MB-231 细胞(HER-,右图)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: HER3/ErbB3 (D22C5) XP® Rabbit mAb Expand Image
使用与浓度匹配的Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (红色)作对比的 HER3/ErbB3 (D22C5) XP® Rabbit mAb (蓝色),对固定和通透的 MCF7 细胞进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor® 488 conjugate) #4412 用作二抗。