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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-GSDMD antibody [EPR20859] (ab219800)

Reviews (2) Submit a question References (35)
Western blot - Anti-GSDMD antibody [EPR20859] (ab219800)
  • Western blot - Anti-GSDMD antibody [EPR20859] (ab219800)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMD antibody [EPR20859] (ab219800)
  • Flow Cytometry (Intracellular) - Anti-GSDMD antibody [EPR20859] (ab219800)
  • Immunoprecipitation - Anti-GSDMD antibody [EPR20859] (ab219800)
  • Anti-GSDMD antibody [EPR20859] (ab219800)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20859] to GSDMD
  • Suitable for: Flow Cyt (Intra), IP, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free PE

Overview

  • Product name

    Anti-GSDMD antibody [EPR20859]
    See all GSDMD primary antibodies

  • Description

    Rabbit monoclonal [EPR20859] to GSDMD

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), IP, WB, IHC-Pmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type mouse liver and lung whole cell lysates; C6, PC-12, RAW 264.7 and NIH/3T3 whole cell lysates; Rat brain and liver lysates. IHC-P: Wild type mouse small intestine tissue. Flow Cyt (intra): RAW 264.7 cells. IP: RAW 264.7 whole cell lysate.

  • General notes

    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 production
    For more information see here.

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

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR20859

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab219800 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt (Intra)
1/500.
IP
1/30.
WB
1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
IHC-P (2)
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
Flow Cyt (Intra)
1/500.
IP
1/30.
WB
1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for ICC/IF.

Target

  • Sequence similarities

    Belongs to the gasdermin family.
  • Information by UniProt

  • Database links

    • Alternative names

      • 1810036L03Rik antibody
      • DF 5L antibody
      • DF5L antibody
      • DFNA 5L antibody
      • DFNA5L antibody
      • FKSG 10 antibody
      • FKSG10 antibody
      • FLJ12150 antibody
      • Gasdermin antibody
      • Gasdermin D antibody
      • Gasdermin domain containing 1 antibody
      • Gasdermin domain containing protein 1 antibody
      • Gasdermin domain-containing protein 1 antibody
      • Gasdermin-D antibody
      • GasderminD antibody
      • GSDMD antibody
      • GSDMD_HUMAN antibody
      • GSDMDC 1 antibody
      • GSDMDC1 antibody
      see all

    Images

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

      Lane 1 : C6 (rat glial tumor cell line) whole cell lysate
      Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
      Lane 3 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
      Lane 4 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
      Lane 5 : Rat liver lysate
      Lane 6 : Rat brain lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : 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.

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

      Lane 1 : Wild-type mouse liver whole cell lysate
      Lane 2 : GSDMD knockout mouse liver whole cell lysate
      Lane 3 : Wild-type mouse lung whole cell lysate
      Lane 4 : GSDMD knockout mouse lung whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : 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, NIBS.

       

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

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

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line labeling GSDMD with ab219800 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Immunoprecipitation - Anti-GSDMD antibody [EPR20859] (ab219800)
      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) (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 (ab172730) instead of ab219800 in RAW 264.7 whole cell lysate.

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

      Exposure time: 10 seconds.

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

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (35)

    Publishing research using ab219800? Please let us know so that we can cite the reference in this datasheet.

    ab219800 has been referenced in 35 publications.

    • Mo G  et al. IP3R1 regulates Ca2+ transport and pyroptosis through the NLRP3/Caspase-1 pathway in myocardial ischemia/reperfusion injury. Cell Death Discov 7:31 (2021). PubMed: 33568649
    • Kim DU  et al. Loganin Attenuates the Severity of Acute Kidney Injury Induced by Cisplatin through the Inhibition of ERK Activation in Mice. Int J Mol Sci 22:N/A (2021). PubMed: 33572597
    • Zou XS  et al. Pomelo peel oil alleviates cerebral NLRP3 inflammasome activation in a cardiopulmonary resuscitation rat model. Exp Ther Med 21:233 (2021). PubMed: 33603841
    • Cao Y  et al. I-?B Kinase-e Deficiency Attenuates the Development of Angiotensin II-Induced Myocardial Hypertrophy in Mice. Oxid Med Cell Longev 2021:6429197 (2021). PubMed: 33628362
    • Zheng X  et al. Dexmedetomidine alleviates myocardial ischemia/reperfusion-induced injury and Ca2+ overload via the microRNA-346-3p/CaMKIId axis. Int J Cardiol 338:185-195 (2021). PubMed: 33731281
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-2 of 2 Abreviews or Q&A

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

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (COLON)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA
    Permeabilization
    No
    Specification
    COLON
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 25 2022

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

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Syrian hamster Cell (Brain tissue)
    Permeabilization
    Yes - 1% Triton X-100 in PBS
    Specification
    Brain tissue
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Apr 21 2021

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