Revision 1
#26776
Store at -20C
Gasdermin Family Antibody Sampler Kit
1 Kit
(6 x 20 microliters)
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes | Product # | Quantity | Mol. Wt | Isotype/Source |
---|---|---|---|---|
Gasdermin D (E5O4N) Rabbit mAb | 69469 | 20 µl | 53, 30 kDa | Rabbit IgG |
Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb | 36425 | 20 µl | 30 kDa | Rabbit IgG |
Gasdermin E (E2X7E) Rabbit mAb | 19453 | 20 µl | 55, 30 kDa | Rabbit IgG |
Cleaved Gasdermin E (Asp270) (E8G4U) Rabbit mAb | 55879 | 20 µl | 30 kDa | Rabbit IgG |
Gasdermin A Antibody | 49307 | 20 µl | 49 kDa | Rabbit |
Gasdermin B Antibody | 76439 | 20 µl | 47 kDa | Rabbit |
Anti-rabbit IgG, HRP-linked Antibody | 7074 | 100 µl | Goat |
Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.
Storage
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).
Like other gasdermin family members, Gasdermin E (also called DFNA5) 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 (8). In addition, cleavage of Gasdermin E can be induced by Granzyme B secreted by NK cells and contributes to tumor suppressive activity (9). Gasdermin E expression is suppressed in several types of cancer, including gastric, colorectal, and breast carcinoma, and may be associated with decreased survival (10-12). In contrast, an increase in Gasdermin E, including the amino-terminal pore-forming fragment, is associated with conditions of excessive inflammation (13-15). Gasdermin A (GSDMA) is preferentially expressed in the epithelium of the skin and gastrointestinal tract and is frequently suppressed in gastric cancer (16-18). Gasdermin B (GSDMB) has been reported to be upregulated in several tumor types, and in breast cancer has been associated with metastasis and poor prognosis (19,20). In addition, Gasdermin B has been associated with immune disorders, including asthma (21,22). Gasdermin B can be cleaved by Granzyme A secreted from cytotoxic lymphocytes leading to pyroptotic cell death (23).
Background References
- Kayagaki, N. et al. (2015) Nature 526, 666-71.
- Shi, J. et al. (2015) Nature 526, 660-5.
- Broz, P. and Dixit, V.M. (2016) Nat Rev Immunol 16, 407-20.
- Aglietti, R.A. et al. (2016) Proc Natl Acad Sci USA 113, 7858-63.
- Ding, J. et al. (2016) Nature 535, 111-6.
- Liu, X. et al. (2016) Nature 535, 153-8.
- Sborgi, L. et al. (2016) EMBO J 35, 1766-78.
- Rogers, C. et al. (2017) Nat Commun 8, 14128.
- Zhang, Z. et al. (2020) Nature 579, 415-420.
- Kim, M.S. et al. (2008) Oncogene 27, 3624-34.
- Kim, M.S. et al. (2008) Biochem Biophys Res Commun 370, 38-43.
- Yokomizo, K. et al. (2012) Anticancer Res 32, 1319-22.
- Tan, G. et al. (2021) Cell Rep 35, 109265.
- Li, Y. et al. (2021) Cell Death Differ 28, 2333-2350.
- Shi, H. et al. (2021) Circ Res 129, 383-396.
- Tamura, M. et al. (2007) Genomics 89, 618-29.
- Saeki, N. et al. (2000) Mamm Genome 11, 718-24.
- Saeki, N. et al. (2007) Oncogene 26, 6488-98.
- Hergueta-Redondo, M. et al. (2014) PLoS One 9, e90099.
- Hergueta-Redondo, M. et al. (2016) Oncotarget 7, 56295-56308.
- Yu, J. et al. (2011) Pediatr Pulmonol 46, 701-8.
- Das, S. et al. (2016) Proc Natl Acad Sci USA 113, 13132-13137.
- Zhou, Z. et al. (2020) Science 368, eaaz7548. doi: 10.1126/science.aaz7548.
Trademarks and Patents
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Revision 1
Western blot analysis of extracts from ACHN cells, untreated or treated with Raptinal (10 μM, indicated times), using Gasdermin E (E2X7E) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from various cell lines using Gasdermin E (E2X7E) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression Gasdermin E protein in MCF7 and Colo 205 cells is consistent with the predicted expression pattern.
Confocal immunofluorescent analysis of ACHN cells (left, positive) and MCF7 cells (right, negative) using Gasdermin E (E2X7E) Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Revision 1
Western blot analysis of extracts from THP-1 cells, differentiated with TPA #4174 (50 ng/ml, overnight) and then treated with LPS #14011 (5 μg/ml, indicated times), using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb (upper), total Gasdermin D (L60) Antibody #93709 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines using Gasdermin A Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). KATO III cells have been shown to lack Gasdermin A expression.
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.
Revision 1
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 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).
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).
Revision 1
Western blot analysis of extracts from control PC-3 cells (lane 1) or CRISPR/Cas9 Gasdermin D knockout (KO) PC-3 cells (lane 2) using Gasdermin D (E5O4N) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from various cell lines using Gasdermin D (E5O4N) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression of Gasdermin D protein in IMR-32 cells is consistent with the predicted expression pattern.
Western blot analysis of extracts from THP-1 cells, differentiated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (50 ng/mL, 24 hr and then rested 48 hr) followed by vehicle (-) or treated with Lipopolysaccharides (LPS) #14011 (5 μg/mL, 6 hr; +), using Gasdermin D (E5O4N) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Revision 1
Immunoprecipitation of Gasdermin D protein from THP-1 extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Rabbit mAb #3900, and lane 3 is Gasdermin D (E5O4N) Rabbit mAb. Western blot analysis was performed using Gasdermin D (E5O4N) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.
Confocal immunofluorescent analysis of control PC-3 cells (left, positive) or CRISPR/Cas9 Gasdermin D knockout (KO) PC-3 cells (right, negative) using Gasdermin D (E5O4N) Rabbit mAb (green), β-Actin (8H10D10) Mouse mAb #3700 (red), or DAPI #4083 (blue).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Revision 1
Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human Gasdermin B (hGSDMB-Myc/DDK; +), using Gasdermin B Antibody.
Immunoprecipitation of Cleaved Gasdermin D (Asp725) from THP-1 cells differentiated with TPA #4174 (50 ng/ml, overnight) and then treated with LPS #14011 (5 μg/ml, 6 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb. Western blot was performed using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation of Gasdermin A from COLO 205 cell extracts. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 is Gasdermin A Antibody. Western blot analysis was performed using Gasdermin A Antibody. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Revision 1
Western blot analysis of extracts from various cell lines using Gasdermin B Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.
Immunoprecipitation of Gasdermin B from COLO 205 extracts. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 Gasdermin B Antibody. Western blot was performed using Gasdermin B Antibody. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Revision 1
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb in the presence of non-cleaved Gasdermin D peptide (left) or Asp275 cleavage-specific Gasdermin D peptide (right).
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's Lymphoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.
Revision 1
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human spleen (left, positive) and skeletal muscle (right, negative) using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded THP-1 cell pellets, differentiated with TPA #4174 (left) and then treated with LPS #14011 (right), using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.
Revision 1
Immunohistochemical analysis of paraffin-embedded human ulcerative colitis using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.