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

    Suitable for: WB, IPmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Synthetic peptide within Mouse GSDMD aa 250-350. The exact sequence is proprietary.
    Database link: Q9D8T2

  • 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

      

    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 rabbit monoclonal antibody.

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

    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

Applications

Our Abpromise guarantee covers the use of ab209845 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
WB 1/1000. Detects a band of approximately 53, 32 kDa (predicted molecular weight: 53 kDa).
IP 1/30.

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

  • 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.

  • 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 (Apoptosis-associated speck-like protein), 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.

    Lane 1: RAW 264.7-derived cells with ectopic expression of ASC whole cell lysate 10ug (Input).

    Lane 2: ab209845 IP in RAW 264.7-derived cells with ectopic expression of ASC whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab209845 in RAW 264.7-derived cells with ectopic expression of ASC whole cell lysate.

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

    Exposure time: 3 minutes.

    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.

  • 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.

  • 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

  • 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.

    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, whole cell lysate 10ug (Input).

    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.

    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.

    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.

References

This product has been referenced in:

  • Zeng QZ  et al. Paclitaxel Enhances the Innate Immunity by Promoting NLRP3 Inflammasome Activation in Macrophages. Front Immunol 10:72 (2019). Read more (PubMed: 30761140) »
  • Tapia VS  et al. The three cytokines IL-1ß, IL-18, and IL-1a share related but distinct secretory routes. J Biol Chem 294:8325-8335 (2019). Read more (PubMed: 30940725) »
See all 13 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Cardiomyocytes)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 31 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Treatment
100 ng/ml LPS in 10% FBS growth meida for 6 h followed by 10 ´M nigericin in 2% FBS starvation media for 2 h
Specification
mouse hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 25 2018

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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