Revision 3
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

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

120, 145

Source/Isotype:

Mouse IgG2b

UniProt ID:

#P11362

Entrez-Gene Id:

2260

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250

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.

Specificity / Sensitivity

Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb detects endogenous levels of FGF receptors only when phosphorylated at tyrosines 653/654. This antibody detects phosphorylated FGF Receptors 2 and 4 when expressed exogenously. Based on sequence comparisons, reactivity with FGF Receptor 3 is possible but has not been experimentally confirmed. The antibody also cross-reacts slightly with activated PDGF and insulin/IGF-I receptors.

Species Reactivity:

Human, Monkey

Species predicted to react based on 100% sequence homology

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr653/654 of human FGF receptor-1. The corresponding sequence is identical in FGF receptor-2, -3 and -4.

Background

Fibroblast growth factors (FGFs) produce mitogenic and angiogenic effects in target cells by signaling through cell surface receptor tyrosine kinases. There are four members of the FGF receptor family: FGFR1 (flg), FGFR2 (bek, KGFR), FGFR3, and FGFR4. Each receptor contains an extracellular ligand-binding domain, a transmembrane domain, and a cytoplasmic kinase domain (1). Following ligand binding and dimerization, the receptors are phosphorylated at specific tyrosine residues (2). Seven tyrosine residues in the cytoplasmic tail of FGFR1 can be phosphorylated: Tyr463, 583, 585, 653, 654, 730, and 766. Tyr653 and Tyr654 are important for catalytic activity of activated FGFR and are essential for signaling (3). The other phosphorylated tyrosine residues may provide docking sites for downstream signaling components, such as Crk and PLCγ (4,5).

  1. Powers, C.J. et al. (2000) Endocr Relat Cancer 7, 165-97.
  2. Reilly, J.F. et al. (2000) J Biol Chem 275, 7771-8.
  3. Mohammadi, M. et al. (1996) Mol Cell Biol 16, 977-89.
  4. Mohammadi, M. et al. (1991) Mol Cell Biol 11, 5068-78.
  5. Larsson, H. et al. (1999) J Biol Chem 274, 25726-34.

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™

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 3
#3476

Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb

Western Blotting Image 1: Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb Expand Image
使用 Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb,对未经处理或经小牛肠磷酸酶 (CIP) 处理的过表达人 FGF 受体-1 的 COS 细胞提取物进行蛋白质印迹分析。人 FGF 受体-1 的过表达导致该受体组成性激活 (致谢加利福尼亚州 Scripps Research Institute 的 Pamela Maher 博士,私人通讯) 。CIP 处理可消除该抗体与 FGF 受体-1 的反应。
Western Blotting Image 2: Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb Expand Image
使用 Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb(上图)和 FGF Receptor 1 (D8E4) XP® Rabbit mAb(下图)对使用或不使用 bFGF 处理的 A-204 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 1: Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb Expand Image
Simple Western️ analysis of lysates (0.01mg/mL) from Cos-7 cells transfected with hFGFR1 using Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb #3476. 虚拟泳道式图像(左图)显示一抗稀释比例在 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 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess Simple Western 仪器上进行该实验。