Revision 1

#39754Store 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):

53, 30

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9D8T2

Entrez-Gene Id:

69146

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

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

Gasdermin D (E9S1X) Rabbit mAb recognizes endogenous levels of total Gasdermin D protein. This antibody recognizes the 30 kDa amino terminal fragment produced during pyroptosis by caspase-1.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu60 of mouse Gasdermin D protein.

Background

Gasdermin D (GSDMD), a member of the gasdermin family that includes GSDMA, GSDMB, and GSDMC, has been reported to have a critical role as a downstream effector of pyroptosis (1,2). Pyroptosis is a lytic type of cell death triggered by inflammasomes, multiprotein complexes assembled in response to pathogen-associated molecular patterns (PAMPs) or danger-associated molecular patterns (DAMPs) that result in the activation of caspase-1 and subsequent cleavage of pro-inflammatory cytokines IL-1β and IL-18 (3). Gasdermin D was identified by two independent groups as a substrate of inflammatory caspases, caspase-1 and caspase-11/4/5, producing two fragments: GSDMD-N and GSDMD-C. Cleavage results in release of an intramolecular inhibitory interaction between the N- and C-terminal domains, allowing the N-terminal fragment GSDMD-N to initiate pyroptosis through the formation of pores on the plasma membrane (4-7).

  1. Kayagaki, N. et al. (2015) Nature 526, 666-71.
  2. Shi, J. et al. (2015) Nature 526, 660-5.
  3. Broz, P. and Dixit, V.M. (2016) Nat Rev Immunol 16, 407-20.
  4. Aglietti, R.A. et al. (2016) Proc Natl Acad Sci U S A 113, 7858-63.
  5. Ding, J. et al. (2016) Nature 535, 111-6.
  6. Liu, X. et al. (2016) Nature 535, 153-8.
  7. Sborgi, L. et al. (2016) EMBO J 35, 1766-78.

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.
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
#39754

Gasdermin D (E9S1X) Rabbit mAb

Western Blotting Image 1: Gasdermin D (E9S1X) Rabbit mAb Expand Image
使用 Gasdermin D (E9S1X) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对对照小鼠骨髓衍生的巨噬细胞(泳道 1)或 Gasdermin D 基因剔除小鼠(泳道 2) mBMDM 的提取物进行蛋白印迹分析。Gasdermin D 基因敲除细胞中无信号证实了抗体对 Gasdermin D 的特异性。来自 Gasdermin D 基因敲除小鼠的 mBMDM 由马萨诸塞州大学伍斯特分校马萨诸塞大学医学院的 Douglas Golenbock 博士提供。
Western Blotting Image 2: Gasdermin D (E9S1X) Rabbit mAb Expand Image
使用 Gasdermin D (E9S1X) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174 (下图)对对照 PC-3 细胞(泳道 1)或 Gasdermin D 敲除型 PC-3 细胞(泳道 2)的提取物进行蛋白印迹分析。Gasdermin D 敲除型 PC-3 细胞中没有信号,这证实了抗体对 Gasdermin D 的特异性。
Western Blotting Image 3: Gasdermin D (E9S1X) Rabbit mAb Expand Image
使用 Gasdermin D (E9S1X) Rabbit mAb(上图)、Cleaved Gasdermin D (Asp276) (E3E3P) Rabbit mAb #10137(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对未经 (-) 或经 Lipopolysaccharides (LPS) #14011(50 ng/ml,4 小时;+)处理后再用 Nigericin (sodium salt) #66419(15 μM,指定时间)处理的小鼠骨髓源性巨噬细胞 (mBMDM) 的提取物进行蛋白印迹分析。
Western Blotting Image 4: Gasdermin D (E9S1X) Rabbit mAb Expand Image
使用 Gasdermin D (E9S1X) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对不同细胞系提取物进行蛋白印迹分析。
Western Blotting Image 5: Gasdermin D (E9S1X) Rabbit mAb Expand Image
使用 Gasdermin D (E9S1X) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对转染空载 (-) 或转染表达带有 Myc/DDK 标签的全长小鼠 Gasdermin D 蛋白 (mGSDMD-Myc/DDK; +) 的表达载体的 293T 细胞提取物进行蛋白印迹分析。
Western Blotting Image 6: Gasdermin D (E9S1X) Rabbit mAb Expand Image
使用 Gasdermin D (E9S1X) Rabbit mAb(上图)、Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb #36425 (中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对通过 TPA (12-O-tetradecanoylphorbol-13-acetate) #4174 分化(50 ng/ml,过夜)然后经过脂多糖 (LPS) #14011 处理 (5 μg/ml,6 小时)的 THP-1 细胞提取物进行蛋白印迹分析。
Immunoprecipitation Image 1: Gasdermin D (E9S1X) Rabbit mAb Expand Image
对 J774A.1 细胞提取物的 Gasdermin D 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 Gasdermin D (E9S1X) Rabbit mAb。使用 Gasdermin D (E9S1X) Rabbit mAb 进行蛋白印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。