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, IP, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

160

Source/Isotype:

Rabbit IgG

UniProt ID:

#P36888

Entrez-Gene Id:

2322

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

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 #48437.

Specificity / Sensitivity

Phospho-FLT3 (Tyr589/591) (E4D6Y) Rabbit mAb recognizes endogenous levels of FLT3 protein only when doubly phosphorylated at Tyr589 and Tyr591.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic double-phospho-peptide corresponding to residues surrounding Tyr589 and Tyr591 of human FLT3 protein.

Background

FMS-related tyrosine kinase 3 (FLT3, also called FLK2) is a member of the Type III receptor tyrosine kinase family, which includes c-Kit, PDGFR, and M-CSF receptors. FLT3 is expressed on early hematopoietic progenitor cells and supports growth and differentiation within the hematopoietic system (1,2). FLT3 is activated after binding with its ligand FL, which results in a cascade of tyrosine autophosphorylation and tyrosine phosphorylation of downstream substrates (3). The p85 subunit of PI3 kinase, SHP2, GRB2, and Shc have all been reported to associate with FLT3 after FL stimulation (4-6). Tyr589/591 is located in the juxtamembrane region of FLT3 and may play an important role in regulation of FLT3 tyrosine kinase activity. Somatic mutations of FLT3 consisting of internal tandem duplications (ITDs) occur in 20% of patients with acute myeloid leukemia (7).

  1. Shurin, M.R. et al. (1998) Cytokine Growth Factor Rev 9, 37-48.
  2. Naoe, T. et al. (2001) Cancer Chemother Pharmacol 48 Suppl 1, S27-30.
  3. Namikawa, R. et al. (1996) Stem Cells 14, 388-95.
  4. Beslu, N. et al. (1996) J Biol Chem 271, 20075-81.
  5. Zhang, S. and Broxmeyer, H.E. (2000) Biochem Biophys Res Commun 277, 195-9.
  6. Zhang, S. et al. (1999) J Leukoc Biol 65, 372-80.
  7. Mizuki, M. et al. (2000) Blood 96, 3907-14.

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 IP: Immunoprecipitation 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 5
#60413

Phospho-FLT3 (Tyr589/591) (E4D6Y) Rabbit mAb

Western Blotting Image 1: Phospho-FLT3 (Tyr589/591) (E4D6Y) Rabbit mAb Expand Image
使用 Phospho-FLT3 (Tyr589/591) (E4D6Y) Rabbit mAb(上图)和 FLT3 (8F2) Rabbit mAb #3462(下图)对经过赋形剂处理 (-) 或已经过 PKC412 #9493 (1 μM,24 小时;+)处理的 SEM 细胞提取物进行蛋白质印迹分析。用 FLT3 激酶抑制剂 PKC412 处理细胞后,对应于磷酸化 FLT3 (Tyr589/591) 的信号丢失表明抗体对磷酸化 FLT3 的特异性。
Immunoprecipitation Image 1: Phospho-FLT3 (Tyr589/591) (E4D6Y) Rabbit mAb Expand Image
对 SEM 细胞提取物磷酸化 FLT3 (Tyr589/591) 蛋白进行免疫沉淀分析。泳道 1 是 10% input,泳道 2 是 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道3是 Phospho-FLT3 (Tyr589/591) (E4D6Y) Rabbit mAb。使用 Phospho-FLT3 (Tyr589/591) (E4D6Y) Rabbit mAb 进行蛋白质印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Immunofluorescence Image 1: Phospho-FLT3 (Tyr589/591) (E4D6Y) Rabbit mAb Expand Image
使用 Phospho-FLT3 (Tyr589/Tyr591) (E4D6Y) Rabbit mAb(绿色)对未经处理(左图)、经过 λ 磷酸酶后处理(中图)或经过 PKC412 #9493(1 μM,24 小时;右图)处理的 SEM 细胞进行共聚焦免疫荧光分析。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Flow Cytometry Image 1: Phospho-FLT3 (Tyr589/591) (E4D6Y) Rabbit mAb Expand Image
使用 Phospho-FLT3 (Tyr589/Tyr591) (E4D6Y) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对经过 PKC412 #9493(1 μM,24 小时;蓝色)或未经处理(蓝色)的 SEM 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。