Revision 5

#71433Store at -20C

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

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

125

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q05397

Entrez-Gene Id:

5747

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
IHC Leica Bond 1:150 - 1:600
Immunohistochemistry (Paraffin) 1:300 - 1:1200
Immunofluorescence (Immunocytochemistry) 1:50 - 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 #60557.

Specificity / Sensitivity

FAK (D5O7U) XP® Rabbit mAb recognizes endogenous levels of total FAK protein. Immunofluorescence was performed on cells plated on tissue culture grade plastic. We did not detect FAK signal on uncoated glass slides.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val793 of human FAK protein.

Background

Focal adhesion kinase (FAK) is a widely expressed cytoplasmic protein tyrosine kinase involved in integrin-mediated signal transduction. It plays an important role in the control of several biological processes, including cell spreading, migration, and survival (1). Activation of FAK by integrin clustering leads to autophosphorylation at Tyr397, which is a binding site for the Src family kinases PI3K and PLCγ (2-5). Recruitment of Src family kinases results in the phosphorylation of Tyr407, Tyr576, and Tyr577 in the catalytic domain, and Tyr871 and Tyr925 in the carboxy-terminal region of FAK (6,7).

  1. Parsons, J.T. et al. (2000) Oncogene 19, 5606-13.
  2. Schaller, M.D. et al. (1994) Mol Cell Biol 14, 1680-8.
  3. Cobb, B.S. et al. (1994) Mol Cell Biol 14, 147-55.
  4. Chen, H.C. et al. (1996) J Biol Chem 271, 26329-34.
  5. Zhang, X. et al. (1999) Proc Natl Acad Sci U S A 96, 9021-6.
  6. Calalb, M.B. et al. (1995) Mol Cell Biol 15, 954-63.
  7. Schlaepfer, D.D. et al. (1994) Nature 372, 786-791.

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)

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.
CST is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered 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 5
#71433

FAK (D5O7U) XP® Rabbit mAb

Western Blotting Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
使用 FAK (D5O7U) XP® Rabbit mAb(上图)或 α-Actinin (D6F6) XP® Rabbit mAb #6487(下图)对不同细胞系提取物进行蛋白质印迹分析。
Western Blotting Image 2: FAK (D5O7U) XP® Rabbit mAb Expand Image
使用 FAK (D5O7U) XP® Rabbit mAb, #71433(上图)或 #8457 β-Actin (D6A8) Rabbit mAb(下图)对对照 HEK293 细胞(泳道 1)或 FAK 敲除型 HEK293 细胞(泳道 2)的提取物进行蛋白印迹分析。FAK 敲除型 HEK293 细胞中没有信号,这证实了抗体对 FAK 的特异性。
Western Blotting Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
使用 FAK (D5O7U) XP® Rabbit mAb #71433 对经过 hEGF(100 ng/mL,30”)处理的 A431 细胞裂解物 (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。
Immunoprecipitation Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
对 RD 细胞裂解物的 FAK 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 FAK (D5O7U) XP® Rabbit mAb。使用 FAK (D5O7U) XP® Rabbit mAb 执行蛋白质印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Immunohistochemistry Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 FAK (D5O7U) XP® Rabbit mAb 对石蜡包埋的人结肠癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: FAK (D5O7U) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 FAK (D5O7U) XP® Rabbit mAb 对石蜡包埋的人非小细胞肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
使用 FAK (D5O7U) XP® Rabbit mAb 对石蜡包埋的人结肠癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: FAK (D5O7U) XP® Rabbit mAb Expand Image
使用 FAK (D5O7U) 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: FAK (D5O7U) XP® Rabbit mAb Expand Image
使用 FAK (D5O7U) XP® Rabbit mAb,对石蜡包埋的人类非霍奇金淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 4: FAK (D5O7U) XP® Rabbit mAb Expand Image
使用 FAK (D5O7U) XP® Rabbit mAb 对石蜡包埋的人卵巢浆液性乳头状癌进行免疫组织化学分析。
Immunohistochemistry Image 5: FAK (D5O7U) XP® Rabbit mAb Expand Image
使用 FAK (D5O7U) XP8226® Rabbit mAb 对石蜡包埋的 HT-29 细胞沉淀物(左图,阳性)和 RPMI 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunofluorescence Image 1: FAK (D5O7U) XP® Rabbit mAb Expand Image
使用 FAK (D5O7U) XP® Rabbit mAb(绿色)对 OVCAR3 细胞(左图,高表达)和 SK-OV-3 细胞(右图,中度表达)进行共聚焦免疫荧光分析。红色 = Propidium Iodide (PI)/RNase Staining Solution #4087。