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Rabbit Recombinant Monoclonal GSDMD antibody. Carrier free. Suitable for IP, WB and reacts with Mouse samples. Cited in 1 publication.

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Images

Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867), expandable thumbnail
  • Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867), expandable thumbnail
  • Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867), expandable thumbnail
  • Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867), expandable thumbnail
  • Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (AB225867), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWB
Mouse
Tested
Tested

Tested
Tested

Species
Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
-
Notes

-

Associated Products

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Target data

Function

Gasdermin-D. Precursor of a pore-forming protein that plays a key role in host defense against pathogen infection and danger signals (PubMed:26375003, PubMed:26375259, PubMed:26611636, PubMed:27383986, PubMed:27385778, PubMed:27418190). This form constitutes the precursor of the pore-forming protein: upon cleavage, the released N-terminal moiety (Gasdermin-D, N-terminal) binds to membranes and forms pores, triggering pyroptosis (PubMed:26375003, PubMed:26375259, PubMed:26611636, PubMed:27383986, PubMed:27385778, PubMed:27418190). Gasdermin-D, N-terminal. Promotes pyroptosis in response to microbial infection and danger signals (PubMed:26375003, PubMed:26375259, PubMed:26611636, PubMed:27383986, PubMed:27385778, PubMed:27418190, PubMed:32820063, PubMed:34289345, PubMed:35705808, PubMed:37988464, PubMed:38530158, PubMed:38538834, PubMed:38632402). Produced by the cleavage of gasdermin-D by inflammatory caspases CASP1 or CASP4/CASP11 in response to canonical, as well as non-canonical (such as cytosolic LPS) inflammasome activators (PubMed:26375003, PubMed:26375259, PubMed:26611636, PubMed:27383986, PubMed:27385778, PubMed:27418190, PubMed:35705808, PubMed:38632402). After cleavage, moves to the plasma membrane where it strongly binds to inner leaflet lipids, including monophosphorylated phosphatidylinositols, such as phosphatidylinositol 4-phosphate, bisphosphorylated phosphatidylinositols, such as phosphatidylinositol (4,5)-bisphosphate, as well as phosphatidylinositol (3,4,5)-bisphosphate, and more weakly to phosphatidic acid and phosphatidylserine (PubMed:27339137, PubMed:27383986). Homooligomerizes within the membrane and forms pores of 10-15 nanometers (nm) of inner diameter, allowing the release of mature interleukin-1 (IL1B and IL18) and triggering pyroptosis (PubMed:27383986, PubMed:29195811, PubMed:29274245, PubMed:33883744, PubMed:38530158, PubMed:38538834). Gasdermin pores also allow the release of mature caspase-7 (CASP7) (PubMed:35705808). In some, but not all, cells types, pyroptosis is followed by pyroptotic cell death, which is caused by downstream activation of ninjurin-1 (NINJ1), which mediates membrane rupture (cytolysis) (PubMed:38632402). Also forms pores in the mitochondrial membrane, resulting in release of mitochondrial DNA (mtDNA) into the cytosol (PubMed:37001519). Gasdermin-D, N-terminal released from pyroptotic cells into the extracellular milieu rapidly binds to and kills both Gram-negative and Gram-positive bacteria, without harming neighboring mammalian cells, as it does not disrupt the plasma membrane from the outside due to lipid-binding specificity (PubMed:27383986). Under cell culture conditions, also active against intracellular bacteria, such as Listeria monocytogenes (PubMed:27383986). Also active in response to MAP3K7/TAK1 inactivation by Yersinia toxin YopJ, which triggers cleavage by CASP8 and subsequent activation (PubMed:30361383, PubMed:30381458). Required for mucosal tissue defense against enteric pathogens (PubMed:37988464). Activation of the non-canonical inflammasome in brain endothelial cells can lead to excessive pyroptosis, leading to blood-brain barrier breakdown (PubMed:38632402). Strongly binds to bacterial and mitochondrial lipids, including cardiolipin. Does not bind to unphosphorylated phosphatidylinositol, phosphatidylethanolamine nor phosphatidylcholine (PubMed:27383986). Gasdermin-D, p13. Transcription coactivator produced by the cleavage by CASP3 or CASP7 in the upper small intestine in response to dietary antigens (PubMed:37327784). Required to maintain food tolerance in small intestine: translocates to the nucleus and acts as a coactivator for STAT1 to induce the transcription of CIITA and MHC class II molecules, which in turn induce type 1 regulatory T (Tr1) cells in upper small intestine (PubMed:37327784). Gasdermin-D, p40. Produced by the cleavage by papain allergen (PubMed:35794369). After cleavage, moves to the plasma membrane and homooligomerizes within the membrane and forms pores of 10-15 nanometers (nm) of inner diameter, allowing the specific release of mature interleukin-33 (IL33), promoting type 2 inflammatory immune response (PubMed:35749514, PubMed:35794369).

Alternative names

Gsdmdc1, Gsdmd, Gasdermin-D, Gasdermin domain-containing protein 1

Recommended products

Rabbit Recombinant Monoclonal GSDMD antibody. Carrier free. Suitable for IP, WB and reacts with Mouse samples. Cited in 1 publication.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR19828
Purification technique
Affinity purification Protein A
Specificity

In our hands this product works well in western blot using mouse cell lines but not tissue samples.

We recommend ab219800 as an alternative for low expression samples.

Expression level of GSDMD in whole normal brain lysate is low or undetectable (PMID: 32671214, PMID: 34975487).

Please navigate to the Downloads section for specificity details in Chinese.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab225867 is the carrier-free version of Anti-GSDMD antibody [EPR19828] ab209845.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

GSDMD also known as gasdermin D is a protein known for its role in pyroptosis a form of programmed cell death. Its molecular weight is approximately 53 kDa. Mechanically GSDMD operates by forming pores in cell membranes. These pores disrupt cellular homeostasis and eventually lead to cell lysis. GSDMD is mainly expressed in immune cells including macrophages and neutrophils. Researchers frequently use GSDMD Western blot and GSDMD ELISA for its detection and quantification in various studies.

Biological function summary

Gasdermin D functions in the execution of immune responses against infections. It acts as an effector molecule that participates directly in pyroptosis by disrupting mitochondrial membranes. GSDMD operates as part of a larger inflammasome complex initiated by inflammatory signals. The inflammasome activates inflammatory caspases that cleave GSDMD enabling its active form to execute pyroptosis. This process releases cytokines like IL-1β enhancing the inflammatory response.

Pathways

GSDMD is important in the pyroptosis pathway initiated by the inflammasome. This process involves Caspase-1 a protease responsible for cleaving pro-inflammatory cytokines and initiating pyroptosis. Another significant pathway includes NLRP3 inflammasome which acts upstream to activate Caspase-1 and subsequently GSDMD establishing the overall inflammatory response in the innate immune system. Through these pathways GSDMD interacts closely with proteins like IL-18 an essential inflammatory mediator.

Associated diseases and disorders

Gasdermin D has links to inflammatory diseases such as rheumatoid arthritis and sepsis. In rheumatoid arthritis the excessive activation of GSDMD leads to chronic joint inflammation mediated by activated immune cells. In sepsis over-activation of the pyroptosis pathway may cause severe systemic inflammation driven by GSDMD activity exacerbating cytokine release. Connections exist between GSDMD and other proteins such as Caspase-11 which can also initiate GSDMD cleavage independently and has roles in non-canonical inflammasome pathways influencing these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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6 product images

  • Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867), expandable thumbnail

    Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)

    Gasdermin-D was immunoprecipitated from 0.35 mg of RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cells with ectopic expression of ASC (Apoptosis-associated speck-like protein), whole cell lysate with Anti-GSDMD antibody [EPR19828] ab209845 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using Anti-GSDMD antibody [EPR19828] ab209845 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.

    Lane 1: RAW 264.7-derived cells with ectopic expression of ASC whole cell lysate 10ug (Input).

    Lane 2: Anti-GSDMD antibody [EPR19828] ab209845 IP in RAW 264.7-derived cells with ectopic expression of ASC whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-GSDMD antibody [EPR19828] ab209845 in RAW 264.7-derived cells with ectopic expression of ASC whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    Details of RAW 264.7-derived cells with ectopic expression of ASC are described in the literature: PMID 26611636.

    The cells were kindly provided by our collaborator Dr. Jiahuai Han, Xiamen University.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GSDMD antibody [EPR19828] ab209845).

    All lanes: Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)

    Predicted band size: 53 kDa

  • Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867), expandable thumbnail

    Immunoprecipitation - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)

    Gasdermin-D was immunoprecipitated from 0.35 mg of RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate with Anti-GSDMD antibody [EPR19828] ab209845 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using Anti-GSDMD antibody [EPR19828] ab209845 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.

    Lane 1: RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate 10ug (Input).

    Lane 2: Anti-GSDMD antibody [EPR19828] ab209845 IP in RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-GSDMD antibody [EPR19828] ab209845 in RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    As a response to LPS stimulation and nigericin treatment, Gasdermin-D is cleaved and Gasdermin-D N-terminal form is detected at 32kDa. Details of RAW 264.7-derived cells with ectopic expression of ASC are described in the literature: PMID 26611636.

    The cells were kindly provided by our collaborator Dr. Jiahuai Han, Xiamen University.

    Note: The antibody has better affinity to full length Gasdermin-D.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GSDMD antibody [EPR19828] ab209845).

    All lanes: Immunoprecipitation - Anti-GSDMD antibody [EPR19828] (Anti-GSDMD antibody [EPR19828] ab209845) at 1/1000 dilution

    Lane 1: RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h at 10 µg

    Lane 2: Anti-GSDMD antibody [EPR19828] ab209845 IP in RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h whole cell lysate at 10 µg

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-GSDMD antibody [EPR19828] ab209845 in RAW 264.7-derived cells with ectopic expression of ASC were primed with 1μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h whole cell lysate

    Predicted band size: 53 kDa

  • Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    As a response to LPS stimulation and nigericin treatment, Gasdermin-D is cleaved and Gasdermin-D N-terminal form is detected at 32kDa. Details of RAW264.7-derived cells with ectopic expression of ASC are described in the literature: PMID 26611636

    The MW observed is consistent with the literature: PMID 26375003.

    The cells were kindly provided by our collaborator Dr. Jiahuai Han, Xiamen University.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GSDMD antibody [EPR19828] ab209845).

    All lanes: Western blot - Anti-GSDMD antibody [EPR19828] (Anti-GSDMD antibody [EPR19828] ab209845) at 1/1000 dilution

    Lane 1: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cell line with ectopic expression of ASC (Apoptosis-associated speck-like protein), whole cell lysate at 20 µg

    Lane 2: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cell line with ectopic expression of ASC primed with 1 μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 μM nigericin treatment under serum starved conditions for 2 h, whole cell lysate at 20 µg

    Lane 3: Gasdermin-D knockout RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus)-derived cell line with ectopic expression of ASC whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 53 kDa

    Observed band size: 32 kDa, 53 kDa

    Exposure time: 1min

  • Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)

    This data was developed using Anti-GSDMD antibody [EPR19828] ab209845, the same antibody clone in a different buffer formulation. Different batches of Anti-GSDMD antibody [EPR19828] ab209845 were tested on:

    1: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) -derived cell line with ectopic expression of ASC primed with 1 μg/mL lipopolysaccharides (LPS) for 4 h followed by 10 uM nigericin treatment under serum starved conditions for 2 h, whole cell lysate at 5.2 μg/ml.

    2: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) -derived cell line with ectopic expression of ASC (Apoptosis-associated speck-like protein), whole cell lysate at 5.2 μg/ml.

    15 μg of lysate was loaded in each lane. Bands observed at 32,53 kDa.

    All lanes: Western blot - Anti-GSDMD antibody [EPR19828] (Anti-GSDMD antibody [EPR19828] ab209845)

    Predicted band size: 53 kDa

  • Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)

    Blocking and dilution buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation (Anti-GSDMD antibody [EPR19828] ab209845).

    All lanes: Western blot - Anti-GSDMD antibody [EPR19828] (Anti-GSDMD antibody [EPR19828] ab209845) at 1/1000 dilution

    Lane 1: J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

    Lane 2: J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) treated with 100ng/ml lipopolysaccharides (LPS) for 3h, then added 300ng/ml BFA for another 3h whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 53 kDa

    Observed band size: 53 kDa

    Exposure time: 180s

  • Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)

    This data was developed using Anti-GSDMD antibody [EPR19828] ab209845, the same antibody clone in a different buffer formulation.

    Compared with Anti-GSDMD antibody [EPR19828] ab209845, Anti-GSDMD antibody [EPR20859] ab219800 has a higher affinity in positive samples.

    The MW observed is consistent with the literature: PMID 26375003.

    The 43 kDa band is likely to be a GSDMD cleavage product by caspase-3 at GSDMD Asp88 (PMID: 30902848, PMID: 28392147, PMID: 30135078).

    We are unsure how to define the 37kDa extra band.

    Other bands may be the GSDMD cleavage products, you can find them in PMID: 34890644, PMID: 33208231, PMID: 28679757.

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-GSDMD antibody [EPR19828] (Anti-GSDMD antibody [EPR19828] ab209845) at 1/1000 dilution

    Lane 1: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate, Batch 1 at 20 µg

    Lane 2: Mouse liver lysate at 20 µg

    Lane 3: NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate at 20 µg

    Lane 4: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate, Batch 2 at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 53 kDa

    Observed band size: 53 kDa

    Exposure time: 120s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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