Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GSDMD antibody [EPR19828] - BSA and Azide free (ab225867)

Overview

  • Product name

    Anti-GSDMD antibody [EPR19828] - BSA and Azide free
    See all GSDMD primary antibodies
  • Description

    Rabbit monoclonal [EPR19828] to GSDMD - BSA and Azide free
  • Host species

    Rabbit
  • 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.
  • General notes

    Ab225867 is the carrier-free version of ab209845. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab225867 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19828
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab225867 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 Use at an assay dependent concentration. Detects a band of approximately 53, 32 kDa (predicted molecular weight: 53 kDa).
IP Use at an assay dependent concentration.

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab209845).

References

ab225867 has not yet been referenced specifically in any publications.

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