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Rabbit Recombinant Monoclonal GSDMD antibody. Suitable for IP, WB and reacts with Mouse samples. Cited in 275 publications.


Images

Immunoprecipitation - Anti-GSDMD antibody [EPR19828] (AB209845), expandable thumbnail
  • Western blot - Anti-GSDMD antibody [EPR19828] (AB209845), expandable thumbnail
  • Western blot - Anti-GSDMD antibody [EPR19828] (AB209845), expandable thumbnail
  • Western blot - Anti-GSDMD antibody [EPR19828] (AB209845), expandable thumbnail
  • Western blot - Anti-GSDMD antibody [EPR19828] (AB209845), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

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

1/30

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

-

Associated Products

Select an associated product type

11 products for Alternative Product

Target data

Function

Gasdermin-DPrecursor 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-terminalPromotes pyroptosis in response to microbial infection and danger signals (PubMed:26375003, PubMed:26375259, PubMed:26611636, PubMed:27383986, PubMed:27385778, PubMed:27418190, PubMed:32820063). 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). 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:27383986, PubMed:27339137). Homooligomerizes within the membrane and forms pores of 10-15 nanometers (nm) of inner diameter, allowing the release of mature IL1B and triggering pyroptosis (PubMed:27383986). Exhibits bactericidal activity (PubMed:27383986). 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). Strongly binds to bacterial and mitochondrial lipids, including cardiolipin. Does not bind to unphosphorylated phosphatidylinositol, phosphatidylethanolamine nor phosphatidylcholine (PubMed:27383986).

Alternative names

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

Recommended products

Rabbit Recombinant Monoclonal GSDMD antibody. Suitable for IP, WB and reacts with Mouse samples. Cited in 275 publications.

Alternative names

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

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

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.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Immunoprecipitation - Anti-GSDMD antibody [EPR19828] (ab209845), expandable thumbnail

    Immunoprecipitation - Anti-GSDMD antibody [EPR19828] (ab209845)

    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 ab209845 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using 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.

    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.

    All lanes: Immunoprecipitation - 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: 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 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] (ab209845), expandable thumbnail

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

    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.

    All lanes: Western blot - 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] (ab209845), expandable thumbnail

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

    Different batches of 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] (ab209845)

    Predicted band size: 53 kDa

  • Western blot - Anti-GSDMD antibody [EPR19828] (ab209845), expandable thumbnail

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

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - 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] (ab209845), expandable thumbnail

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

    Compared with 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] (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

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