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Rabbit Recombinant Monoclonal GSDMD antibody. Carrier free. Suitable for IP, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Rat samples. Cited in 3 publications.

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Images

Western blot - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (AB239377), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (AB239377), expandable thumbnail
  • Immunoprecipitation - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (AB239377), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (AB239377), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (AB239377), 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
IPWBFlow Cyt (Intra)IHC-P
Mouse
Tested
Tested
Tested
Tested
Rat
Predicted
Expected
Predicted
Predicted

Tested
Tested

Species

Mouse

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Predicted
Predicted

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

-

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

-

Notes

-

Predicted
Predicted

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Predicted
Predicted

Species

Rat

Dilution info

-

Notes

-

Associated Products

Select an associated product type

9 products for Alternative Product

2 products for Alternative Version

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, 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, p13Transcription 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, p40Produced 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, Gsdmdc1, Gasdermin-D, Gasdermin domain-containing protein 1

Recommended products

Rabbit Recombinant Monoclonal GSDMD antibody. Carrier free. Suitable for IP, WB, Flow Cyt (Intra), IHC-P and reacts with Mouse, Rat samples. Cited in 3 publications.

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

EPR20859

Purification technique

Affinity purification Protein A

Specificity

This antibody can detect full length, as well as a N-terminal fragment after stimulation in WB.

Expression level of GSDMD in whole normal brain lysate is low or undetectable (PMID: 32671214, PMID: 34975487), we recommend loading higher amount of lysate or using lower antibody dilution.

Please see documentation for further information on positive controls (Chinese version)

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

ab239377 is the carrier-free version of Anti-GSDMD antibody [EPR20859] ab219800.

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.

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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8 product images

  • Western blot - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377)

    Exposure time : Lanes 1 and 2:3 minutes; Lanes 3 and 4:48 seconds; Lanes 5 and 6:3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Expression level of GSDMD in whole normal brain lysate is low or undetectable (PMID: 32671214, PMID: 34975487), we recommend loading higher amount of lysate or using lower antibody dilution.



    This data was developed using Anti-GSDMD antibody [EPR20859] ab219800, the same antibody clone in a different buffer formulation.

    All lanes: Western blot - Anti-GSDMD antibody [EPR20859] (Anti-GSDMD antibody [EPR20859] ab219800) at 1/1000 dilution

    Lane 1: C6 (rat glial tumor cell line) whole cell lysate at 10 µg

    Lane 2: PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

    Lane 3: RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

    Lane 4: NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate at 10 µg

    Lane 5: Rat liver lysate at 10 µg

    Lane 6: Rat brain lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 53 kDa

    Observed band size: 53 kDa

    Exposure time : Lanes 1 and 2:3 minutes; Lanes 3 and 4:48 seconds; Lanes 5 and 6:3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Expression level of GSDMD in whole normal brain lysate is low or undetectable (PMID: 32671214, PMID: 34975487), we recommend loading higher amount of lysate or using lower antibody dilution.

  • Flow Cytometry (Intracellular) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377)

    Flow cytometry overlay histogram showing Raw264.7 cells stained with Anti-GSDMD antibody [EPR20859] ab219800 (red line). The cells were fixed with 80% Methanol and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10µg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (Anti-GSDMD antibody [EPR20859] ab219800) (1x 106 in 100µl at 0.2µg/ml (1/2)) for 30min at 22°C.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C.

    Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

  • Immunoprecipitation - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377), expandable thumbnail

    Immunoprecipitation - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377)

    GSDMD was immunoprecipitated from 0.35 mg of RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate with Anti-GSDMD antibody [EPR20859] ab219800 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-GSDMD antibody [EPR20859] ab219800 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.

    Lane 1: RAW 264.7 whole cell lysate 10 μg (Input).

    Lane 2: Anti-GSDMD antibody [EPR20859] ab219800 IP in RAW 264.7 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-GSDMD antibody [EPR20859] ab219800 in RAW 264.7 whole cell lysate.

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

    Exposure time: 10 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GSDMD antibody [EPR20859] ab219800).

    All lanes: Immunoprecipitation - Anti-GSDMD antibody [EPR20859] (Anti-GSDMD antibody [EPR20859] ab219800)

    Predicted band size: 53 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377)

    Immunohistochemical analysis of paraffin-embedded Wild type mouse small intestine (A) and GSDMD KO mouse small intestine (B) tissue labeling GSDMD with Anti-GSDMD antibody [EPR20859] ab219800 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on epithelium of wild type mouse small intestine (A), no staining on GSDMD KO mouse small intestine (B) (PMID: 17350798) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GSDMD antibody [EPR20859] ab219800).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    The tissue samples were kindly provided by Dr Feng Shao's lab, NIBS.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377)

    Immunohistochemical analysis of paraffin-embedded mouse lung tissue labelling GSDMD with Anti-GSDMD antibody [EPR20859] ab219800 at 1/1000 (0.542 μg/ml) followed by Goat Anti-Rabbit IgG H&L (HRP polymer) ab214880 at a ready to use dilution. Low expression tissue: weak staining on mouse lung. The section was incubated with Anti-GSDMD antibody [EPR20859] ab219800 at 4°C overnight. Counterstained with hematoxylin.

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (HRP polymer) ab214880 at a ready to use dilution.

    Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GSDMD antibody [EPR20859] ab219800).

  • Western blot - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GSDMD antibody [EPR20859] ab219800).

    Blocking and diluting buffer: 5% NFDM /TBST

    This antibody could detect full length, as well as a N-terminal fragment after stimulation.

    The lysates were kindly provided by Dr Feng Shao's lab, NIBS.

    All lanes: Western blot - Anti-GSDMD antibody [EPR20859] (Anti-GSDMD antibody [EPR20859] ab219800) at 1/2000 dilution

    Lane 1: iBMM (mouse immortalized bone marrow derived macrophages) treated with 500ng/ml Bsak plus 500ng/ml anthrax protective antigen (PA) for 2h. Cell lysate

    Lane 2: iBMM (mouse immortalized bone marrow derived macrophages) treated with 500ng/ml Bsak plus 500ng/ml anthrax protective antigen (PA) for 2h. Concentrated cell supernatant

    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, 31 kDa

    Observed band size: 31 kDa, 53 kDa

    Blocking and diluting buffer: 5% NFDM /TBST

    This antibody could detect full length, as well as a N-terminal fragment after stimulation.

    The lysates were kindly provided by Dr Feng Shao's lab, NIBS.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377)

    This data was developed using Anti-GSDMD antibody [EPR20859] ab219800, the same antibody clone in a different buffer formulation.
    Immunohistochemical analysis of paraffin-embedded (A) Small intestine tissue from wild-type C57BL/6JGpt mice (B) Small intestine tissue from GSDMD knockout mice staining with Anti-GSDMD antibody [EPR20859] ab219800 at 1/5000 dilution and ready-to-use Goat Anti-Rabbit IgG H&L (HRP) secondary. Counterstaining with hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Positive staining on (A) Small intestine tissue from wild-type C57BL/6JGpt mice and no staining on (B) Small intestine tissue from GSDMD knockout mice. The section was incubated with Anti-GSDMD antibody [EPR20859] ab219800 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND™ RX instrument. The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and GSDMD-KO homozygous mice (Strain ID: T010437).

  • Western blot - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR20859] - BSA and Azide free (ab239377)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GSDMD antibody [EPR20859] ab219800).

    Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] ab215203 mainly recognizes N-terminal GSDMD and Anti-GSDMD antibody [EPR20859] ab219800 mainly recognizes full length GSDMD.

    Exposure time: Lanes 1-4: 100 seconds, Lanes 5-6: 40 seconds.

    Lanes 1 - 4: Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] ab215203) at 1/1000 dilution

    Lanes 5 - 6: Western blot - Anti-GSDMD antibody [EPR20859] (Anti-GSDMD antibody [EPR20859] ab219800) at 1/1000 dilution

    Lanes 1, 3 and 5: Untreated THP-1 (human epidermoid carcinoma cell line) whole cell lysate at 20 µg

    Lanes 2, 4 and 6: THP-1 (human epidermoid carcinoma cell line) treated with 50ng/ml TPA for 24 hours, then treated with 5ng/ml LPS for 3 hours and add 1µg/ml BFA for another 3 h whole cell lysate at 20 µg

    Secondary

    Lanes 1, 2, 5 and 6: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Lanes 3 - 4: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/5000 dilution

    Observed band size: 31 kDa

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