Revision 1
Cell Signaling Technology

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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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#55879

Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb

Western Blotting Image 1: Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb Expand Image
Western blot analysis of extracts from ACHN and MCF7 cells, untreated (-) or treated with Raptinal (10 μM, 2 hr; +), using Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb (upper), Gasdermin E (E2X7E) Rabbit mAb #19453 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression of Gasdermin E protein in MCF7 cells is consistent with the predicted expression pattern.
Western Blotting Image 2: Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb Expand Image
Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 4 hr; +), using Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb (upper), Gasdermin E Antibody #84005 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 3: Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb Expand Image
Western blot analysis of extracts from ACHN cells, untreated (-) or treated with Raptinal (10 μM, indicated times), using Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb (upper), Gasdermin E Antibody #84005 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunofluorescence Image 1: Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of ACHN cells, either untreated (left) or treated with Raptinal (10 µM, 1 hr; middle), and MCF7 cells treated with Raptinal (10 µM, 1 hr; right) using Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb (green) and DAPI #4083 (blue).