Revision 1

#50542Store 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, IP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

22-26

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q96LC9

Entrez-Gene Id:

90427

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 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.

Specificity / Sensitivity

BMF (E5U2J) Rabbit mAb recognizes endogenous levels of total BMF protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu47 of human BMF protein.

Background

The BH3-only proteins are a group of pro-apoptotic proteins of the Bcl-2 family that share the conserved BH3 domain but lack BH1, BH2, and BH4 (1). This short BH3 domain is essential for interaction with pro-survival members of the Bcl-2 family and allows for their pro-apoptotic activity. A large number of BH3-only proteins have been identified in mammals, including Bmf, Bad, Bik, Bid, Bim, Hrk, Noxa, and Puma. Many of these proteins appear to display distinct roles in apoptosis through tissue-specific expression. Bmf (Bcl-2-modifying factor) was originally identified in a yeast two-hybrid screen using the pro-survival protein Mcl-1 as bait (2). Bmf appears to be widely expressed, with bmf mRNA observed in cell lines of B- and T-lymphoid, myeloid, or fibroblastoid origin and in mouse embryos at all developmental stages. Bmf protein is seen most abundantly in pancreas, liver, kidney, and hematopoietic tissues (2, 3). Bmf interacts with several pro-survival Bcl-2 proteins including Mcl-1, Bcl-2, Bcl-xL, and Bcl-w, and the interaction depends on the BH3 domain (2). Like Bim, Bmf has been reported to bind to cytoskeletal structures. Bmf is normally sequestered to myosin V motors through association with dynein light chain 2 (DLC2). Certain damage signals, such as the detachment of adherent cell lines from their substratum (anoikis), triggers the release of Bmf and subsequent binding to the pro-survival Bcl-2 proteins (2).

  1. Puthalakath, H. and Strasser, A. (2002) Cell Death Differ 9, 505-12.
  2. Puthalakath, H. et al. (2001) Science 293, 1829-32.
  3. Morales, A.A. et al. (2004) Leukemia 18, 41-7.

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 IP: Immunoprecipitation

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.
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 1
#50542

BMF (E5U2J) Rabbit mAb

Western Blotting Image 1: BMF (E5U2J) Rabbit mAb Expand Image
使用 BMF (E5U2J) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对不同细胞系的提取物进行蛋白质印迹分析。根据 RNAseq 数据预测 NCI-H226 细胞中的信号减低,并确认抗体对 BMF 的特异性。
Western Blotting Image 2: BMF (E5U2J) Rabbit mAb Expand Image
使用 BMF (E5U2J) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对转染空载或转染带 BMF siRNA(48 小时;+)的 A-375 细胞提取物进行蛋白质印迹分析。转染后细胞未经处理(-)或经过 vemurafenib(10μM,18小时;+)的处理。
Western Blotting Image 3: BMF (E5U2J) Rabbit mAb Expand Image
使用 BMF (E5U2J) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5147(下图)对转染空载 (-) 或转染表达带有 Myc/DDK 标签的全长人 BMF 蛋白 (hBMF-Myc/DDK; +) 的表达载体的 293T 细胞的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: BMF (E5U2J) Rabbit mAb Expand Image
对 NCI-H209 细胞提取物 BMF 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 BMF (E5U2J) Rabbit mAb。使用 BMF (E5U2J) Rabbit mAb 进行蛋白质印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。