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Anti-GSDMD antibody [EPR20859] ab219800 is a rabbit monoclonal antibody that is used in GSDMD western blotting, IHC and flow cytometry. Suitable for mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with GSDMD knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR20859 is cited in over 240 publications


Images

Western blot - Anti-GSDMD antibody [EPR20859] (AB219800), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-GSDMD antibody [EPR20859] (AB219800), expandable thumbnail
  • Western blot - Anti-GSDMD antibody [EPR20859] (AB219800), expandable thumbnail
  • Immunoprecipitation - Anti-GSDMD antibody [EPR20859] (AB219800), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] (AB219800), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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
IPWBICC/IFFlow Cyt (Intra)IHC-P
Mouse
Tested
Tested
Not recommended
Tested
Tested
Rat
Expected
Tested
Not recommended
Expected
Expected

Tested
Tested

Species

Mouse

Dilution info

1/30

Notes

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

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1999

Notes

-

Species

Rat

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/500

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

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

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

9 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

Anti-GSDMD antibody [EPR20859] ab219800 is a rabbit monoclonal antibody that is used in GSDMD western blotting, IHC and flow cytometry. Suitable for mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with GSDMD knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR20859 is cited in over 240 publications

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
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 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:
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11 product images

  • Western blot - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR20859] (ab219800)

    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.

    All lanes: Western blot - 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

  • Flow Cytometry (Intracellular) - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-GSDMD antibody [EPR20859] (ab219800)

    Flow cytometry overlay histogram showing Raw264.7 cells stained with 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 (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.

  • Western blot - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR20859] (ab219800)

    Exposure time: Lanes 1 and 2:15 seconds; Lanes 3 and 4:37 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The antibody has strong reactivity with GSDMD with a few weak bands of undermined proteins.

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

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

    Lane 1: Wild-type mouse liver whole cell lysate at 20 µg

    Lane 2: GSDMD knockout mouse liver whole cell lysate at 20 µg

    Lane 3: Wild-type mouse lung whole cell lysate at 20 µg

    Lane 4: GSDMD knockout mouse lung 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 1/100000 dilution

    Predicted band size: 53 kDa

    Observed band size: 53 kDa

    Exposure time: Lanes 1 and 2:15 seconds; Lanes 3 and 4:37 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The antibody has strong reactivity with GSDMD with a few weak bands of undermined proteins.

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

  • Immunoprecipitation - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Immunoprecipitation - Anti-GSDMD antibody [EPR20859] (ab219800)

    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 ab219800 at 1/30 dilution. Western blot was performed from the immunoprecipitate using 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: ab219800 IP in RAW 264.7 whole cell lysate.

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

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

    Exposure time: 10 seconds.

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

    Predicted band size: 53 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] (ab219800)

    Immunohistochemical analysis of paraffin-embedded Wild type mouse small intestine (A) and GSDMD KO mouse small intestine (B) tissue labeling GSDMD with 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.

    The tissue samples were kindly provided by our collaborator Dr. Feng Shao NIBS.

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

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

  • Western blot - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Western blot - Anti-GSDMD antibody [EPR20859] (ab219800)

    Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] ab215203 mainly recognizes N-terminal GSDMD and 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] (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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] (ab219800)

    Immunohistochemical analysis of paraffin-embedded mouse lung tissue labelling GSDMD with 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 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).

  • Immunohistochemistry - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Immunohistochemistry - Anti-GSDMD antibody [EPR20859] (ab219800)

    Immunohistochemical analysis of formalin fixed paraffin embedded mouse small intestine labelling GSDMD with ab219800 at a concentration of 0.1µ/ml. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20mins.

    ab219800 anti-GSDMD antibody [EPR20859] was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.

    Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

  • Immunohistochemistry - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Immunohistochemistry - Anti-GSDMD antibody [EPR20859] (ab219800)

    Immunohistochemical analysis of formalin fixed paraffin embedded mouse small intestine labelling GSDMD with ab219800 at a concentration of 0.05µ/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

    ab219800 anti-GSDMD antibody [EPR20859] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

    Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

  • Western blot - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Western blot - 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] (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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] (ab219800), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] (ab219800)

    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 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 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).

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