Revision 1

#56343Store 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):

55, 25

Source/Isotype:

Rabbit IgG

UniProt ID:

#O15519

Entrez-Gene Id:

8837

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 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.

Specificity / Sensitivity

FLIP (D5J1E) Rabbit mAb recognizes endogenous levels of total FLIP protein. This antibody recognizes both the long and short isoforms of FLIP.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human FLIP protein.

Background

Cellular FLIP (FLICE inhibitory protein) is a regulator of apoptosis that has various names, such as c-FLIP (1), Casper (2), CLARP (3), FLAME (4), I-FLICE (5), MRIT (6), CASH (7), and Usurpin (8). FLIP is expressed as two alternative splice isoforms, FLIP short (FLIPS) and FLIP long (FLIPL). FLIPS contains two death effector domains (DEDs) like those found on the death receptor adaptor protein FADD and the pro-domain of caspase-8. FLIPL shares significant homology with caspase-8 (FLICE), and contains an additional death effector domain, but FLIPL lacks the catalytic active site of the caspases and does not have protease activity. Both FLIP isoforms have been reported to interact with FADD and pro-caspase-8. The role of FLIP in apoptosis is controversial as some research studies have reported it to be anti-apoptotic, while others claim that it is pro-apoptotic. Overexpression of FLIPL can lead to caspase-8 heterodimers that produce an active protease, resulting in apoptosis. However, at physiological levels, it is thought that the binding of FLIP to the DED of FADD results in inhibition of caspase-8 processing. Reduction of FLIP by siRNA or gene targeting sensitizes cells to death receptor-mediated apoptosis. FLIP has also been implicated in the resistance of cancer cells to apoptosis and is upregulated in some cancer types including Hodgkin's lymphoma and ovarian and colon carcinomas (9).

  1. Irmler, M. et al. (1997) Nature 388, 190-5.
  2. Shu, H.B. et al. (1997) Immunity 6, 751-63.
  3. Inohara, N. et al. (1997) Proc Natl Acad Sci U S A 94, 10717-22.
  4. Srinivasula, S.M. et al. (1997) J Biol Chem 272, 18542-5.
  5. Hu, S. et al. (1997) J Biol Chem 272, 17255-7.
  6. Han, D.K. et al. (1997) Proc Natl Acad Sci U S A 94, 11333-8.
  7. Rasper, D.M. et al. (1998) Cell Death Differ 5, 271-88.
  8. Goltsev, Y.V. et al. (1997) J Biol Chem 272, 19641-4.
  9. Kataoka, T. (2005) Crit Rev Immunol 25, 31-58.

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

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.
SignalSilence 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 1
#56343

FLIP (D5J1E) Rabbit mAb

Western Blotting Image 1: FLIP (D5J1E) Rabbit mAb Expand Image
使用 FLIP (D5J1E) Rabbit mAb(上)或 β-Actin (D6A8) Rabbit mAb #8457(下)对不同细胞系提取物进行蛋白质印迹分析。KARPAS 细胞系来源:剑桥大学 Abraham Karpas 博士。
Western Blotting Image 2: FLIP (D5J1E) Rabbit mAb Expand Image
使用 FLIP (D5J1E) Rabbit mAb 对转染空载 (-) 或经构建表达全长人 FLIP (hFLIP, +) 转染的 293T 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 3: FLIP (D5J1E) Rabbit mAb Expand Image
使用 FLIP (D5J1E) Rabbit mAb (上)或 β-Actin (D6A8) #8457(下),对经 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-)、SignalSilence® FLIP siRNA I #12372 (+) 或 SignalSilence® FLIP siRNA II #12407 (+) 转染的 A549 细胞提取物进行蛋白质印迹分析。FLIP (D5J1E) Rabbit mAb 确认了 FLIP 表达的沉默,而 β-Actin (D6A8) Rabbit mAb 则被用于上样量对照。
Immunoprecipitation Image 1: FLIP (D5J1E) Rabbit mAb Expand Image
对 KARPAS-299 细胞提取物中的 FLIP 进行免疫沉淀。泳道 1 是 10% 的输入,泳道 2 则是经 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 沉淀的,而泳道 3 是 FLIP (D5J1E) Rabbit mAb。使用 FLIP (D5J1E) Rabbit mAb 进行蛋白质印迹分析。为避免与 IgG 的交叉反应性,需使用具有构象特异性的二抗。KARPAS 细胞系来源:剑桥大学 Abraham Karpas 博士。