Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-GSDMD antibody [EPR19828] (ab209845)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19828] to GSDMD
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse

Overview

  • Product name

    Anti-GSDMD antibody [EPR19828]
    See all GSDMD primary antibodies

  • Description

    Rabbit monoclonal [EPR19828] to GSDMD

  • Host species

    Rabbit

  • 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 western blotting of mouse tissue samples.

  • Tested Applications & Species

    Application Species
    IP
    Mouse
    WB
    Mouse
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: RAW 264.7-derived cell line with ectopic expression of ASC (Apoptosis-associated speck-like protein), untreated and 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 lysates; J774A.1 IP: RAW 264.7-derived cells with ectopic expression of ASC, untreated and 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 lysates.

  • 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: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR19828

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)
    Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)
    Lanes 1-3 : Anti-GSDMD antibody [EPR19828] (ab209845) at 1/1000 dilution
    Lanes 4-6 : Anti-GSDMD antibody [EPR20859] (ab219800) at 1/1000 dilution

    Lanes 1 & 4 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
    Lanes 2 & 5 : Mouse liver tissue lysate
    Lanes 3 & 6 : Mouse spinal cord tissue lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 53 kDa
    Observed band size: 53 kDa


    Exposure time: 3 minutes


    Blocking/Diluting buffer and concentration 5% NFDM/TBST.

    Ab209845 work best with cell lysates.

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

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 53 kDa
    Observed band size: 32,53 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    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.

  • Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)
    Western blot - Anti-GSDMD antibody [EPR19828] (ab209845)
    Lane 1 : Anti-GSDMD antibody [EPR19828] (ab209845) at 1/1000 dilution
    Lane 2 : Anti-GSDMD antibody [EPR20859] (ab219800) at 1/1000 dilution

    All lanes : J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) whole cell lysates

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 53 kDa
    Observed band size: 53 kDa


    Exposure time: 3 minutes


    Diluting/Blocking buffer and concentration 5% NFDM/TBST

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

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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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