Revision 1
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, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

30

Source/Isotype:

Rabbit IgG

UniProt ID:

#O60443

Entrez-Gene Id:

1687

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 1:100 - 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

Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb recognizes endogenous levels of Gasdermin E protein only when cleaved at Asp270. A band of unknown origin is detected at around 25 kDa.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp270 of human Gasdermin E protein.

Background

The gasdermin family, which includes GSDMA, GSDMB, GSDMC, GSDMD, and GSDME, has been shown to play a role in inflammation and cell death. Gasdermin D 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).

Gasdermin E (GSDME), also known as DFNA5, was originally identified as a genetic cause of nonsyndromic hearing loss (8). Like other gasdermin family members, Gasdermin E contains an amino-terminal pore forming domain that triggers pyroptosis. Cleavage of Gasdermin E at Asp270 is induced by apoptotic-associated Caspase-3, converting apoptotic signals to pyroptosis (9). In addition, cleavage of Gasdermin E can be induced by Granzyme B secreted by NK cells and contributes to tumor suppressive activity (10). Gasdermin E expression is suppressed in several types of cancer including gastric, colorectal, and breast carcinoma, and may be associated with decreased survival (11-13). In contrast, an increase in Gasdermin E, including the amino-terminal pore-forming fragment, is associated with conditions of excessive inflammation (14-16).

  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 USA 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.
  8. Van Laer, L. et al. (1998) Nat Genet 20, 194-7.
  9. Rogers, C. et al. (2017) Nat Commun 8, 14128.
  10. Zhang, Z. et al. (2020) Nature 579, 415-420.
  11. Kim, M.S. et al. (2008) Oncogene 27, 3624-34.
  12. Kim, M.S. et al. (2008) Biochem Biophys Res Commun 370, 38-43.
  13. Yokomizo, K. et al. (2012) Anticancer Res 32, 1319-22.
  14. Tan, G. et al. (2021) Cell Rep 35, 109265.
  15. Li, Y. et al. (2021) Cell Death Differ 28, 2333-2350.
  16. Shi, H. et al. (2021) Circ Res 129, 383-396.

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 IF-IC: Immunofluorescence (Immunocytochemistry)

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

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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
#55879

Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb

Western Blotting Image 1: Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb Expand Image
使用 Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb(上图)、Gasdermin E (E2X7E) Rabbit mAb #19453(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对未经处理 (-) 或已经过 Raptinal(10μM,2 小时;+)处理的 ACHN 和 MCF77细胞的提取物进行蛋白质印迹法分析。Gasdermin E 蛋白在 MCF7 细胞中的阴性表达与预测的表达模式一致。
Western Blotting Image 2: Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb Expand Image
使用 Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb(上图)、Gasdermin E Antibody #84005(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对未经处理 (-) 或经过 Staurosporine #9953(1 μM,4 小时;+)处理的 Hela 细胞的提取物进行蛋白质印迹法分析。
Western Blotting Image 3: Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb Expand Image
使用 Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb(上图)、Gasdermin E Antibody #84005(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图),对未经处理 (-) 或经过 Raptinal(10 μM,指定时间)处理的 ACHN 细胞的提取物进行蛋白质印迹法分析。
Immunofluorescence Image 1: Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb Expand Image
使用 Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb(绿色)和 DAPI #4083(蓝色),对未经处理(左图)或经过 Raptinal(10 µM,1 小时;中图)处理的 ACHN 细胞和经过 Raptinal(10 µM,1 小时;右图)处理的 MCF7 细胞进行共聚焦免疫荧光分析。