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RecombinantRabMAb

Recombinant Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (3) Submit a question References (26)

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Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)
  • Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)
  • Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20829-408] to cleaved N-terminal GSDMD
  • Suitable for: WB
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-cleaved N-terminal GSDMD antibody [EPR20829-408]
    See all cleaved N-terminal GSDMD primary antibodies
  • Description

    Rabbit monoclonal [EPR20829-408] to cleaved N-terminal GSDMD
  • Host species

    Rabbit
  • Specificity

    The N-term cleaved Gasdermin-D needs inflammatory caspases in response to canonical and non-canonical inflammasome activators (PubMed: 31548300, PubMed: 27418190, PubMed: 26375259). If need to detect cleavage of GSDMD, please ensure that the samples are treated with inflammation before testing.

  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: Salmonella Infected (MOI:100) A431 whole cell lysate; THP-1 treated with 500 ng/ml EprI for 2h, 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 (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20829-408
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Other Antibodies
    • Other Antibodies

Associated products

  • Alternative Versions

    • Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] - BSA and Azide free (ab255983)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Related Products

    • Anti-GSDMD antibody [EPR19829] (ab210070)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab215203 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)
1/1000. Detects a band of approximately 31 kDa (predicted molecular weight: 53 kDa).
Notes
WB
1/1000. Detects a band of approximately 31 kDa (predicted molecular weight: 53 kDa).
Application notes
Is unsuitable for IHC-P.

Target

  • Function

    Gasdermin-D, N-terminal: Promotes pyroptosis in response to microbial infection and danger signals. Produced by the cleavage of gasdermin-D by inflammatory caspases CASP1 or CASP4 in response to canonical, as well as non-canonical (such as cytosolic LPS) inflammasome activators (PubMed:26375003, PubMed:26375259, 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:27281216). Homooligomerizes within the membrane and forms pores of 10 - 15 nanometers (nm) of inner diameter, possibly allowing the release of mature IL1B and triggering pyroptosis (PubMed:27418190, PubMed:27281216). Exhibits bactericidal activity. 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:27281216). Under cell culture conditions, also active against intracellular bacteria, such as Listeria monocytogenes (By similarity). Strongly binds to bacterial and mitochondrial lipids, including cardiolipin. Does not bind to unphosphorylated phosphatidylinositol, phosphatidylethanolamine nor phosphatidylcholine (PubMed:27281216).
  • Tissue specificity

    Expressed in the suprabasal cells of esophagus, as well as in the isthmus/neck, pit, and gland of the stomach, suggesting preferential expression in differentiating cells.
  • Sequence similarities

    Belongs to the gasdermin family.
  • Domain

    Intramolecular interactions between N- and C-terminal domains may be important for autoinhibition in the absence of cleavage by inflammatory caspases CASP1 or CASP4. The intrinsic pyroptosis-inducing activity is carried by gasdermin-D, N-terminal, that is released upon cleavage by inflammatory caspases.
  • Post-translational
    modifications

    Cleavage at Asp-275 by CASP1 (mature and uncleaved precursor forms) or CASP4 relieves autoinhibition and is sufficient to initiate pyroptosis.
  • Cellular localization

    Cell membrane. Secreted. Released in the extracellular milieu following pyroptosis and Cytoplasm, cytosol. Inflammasome. In response to a canonical inflammasome stimulus, such as nigericin, recruited to NLRP3 inflammasone with similar kinetics to that of uncleaved CASP1 precursor.
  • Target information above from: UniProt accession P57764 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 79792 Human
    • SwissProt: P57764 Human
    • Unigene: 118983 Human
    • Alternative names

      • C-terminal antibody
      • Gasdermin domain-containing protein 1 antibody
      • Gasdermin-D antibody
      • GSDMD antibody
      • GSDMD-CT antibody
      • GSDMD-NT antibody
      • GSDMD_HUMAN antibody
      see all

    Images

    • Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)
      Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)
      Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203) at 1/1000 dilution + Salmonella Infected (MOI:100) A431 (human epidermoid carcinoma cell line) whole cell lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 53 kDa
      Observed band size: 31 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Lysate was kindly provided by Dr Feng Shao.

      The molecular weight observed is consistent with literature (PMID: 26375003).

      Blocking/Dilution buffer: 5% NFDM/TBST.

      The N-term cleaved Gasdermin-D needs inflammatory caspases in response to canonical and non-canonical inflammasome activators (PubMed: 31548300, PubMed: 27418190, PubMed: 26375259). If need to detect cleavage of GSDMD, please ensure that the samples are treated with inflammation before testing.

    • Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)
      Western blot - Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)
      All lanes : Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203) at 1/1000 dilution

      Lane 1 : Untreated THP-1 (human epidermoid carcinoma cell line) whole cell lysate
      Lane 2 : THP-1 treated with 500 ng/ml EprI for 2h, 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: 31 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Lysate was kindly provided by Dr Feng Shao.

      The molecular weight observed is consistent with literature (PMID: 26375003).

      Blocking/Dilution buffer: 5% NFDM/TBST.

      The N-term cleaved Gasdermin-D needs inflammatory caspases in response to canonical and non-canonical inflammasome activators (PubMed: 31548300, PubMed: 27418190, PubMed: 26375259). If need to detect cleavage of GSDMD, please ensure that the samples are treated with inflammation before testing.

    • Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)
      Anti-cleaved N-terminal GSDMD antibody [EPR20829-408] (ab215203)

    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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (26)

    Publishing research using ab215203? Please let us know so that we can cite the reference in this datasheet.

    ab215203 has been referenced in 26 publications.

    • Yu X  et al. Long noncoding RNA MIAT regulates primary human retinal pericyte pyroptosis by modulating miR-342-3p targeting of CASP1 in diabetic retinopathy. Exp Eye Res 202:108300 (2021). PubMed: 33065089
    • Zhao K  et al. Acid-sensing ion channels regulate nucleus pulposus cell inflammation and pyroptosis via the NLRP3 inflammasome in intervertebral disc degeneration. Cell Prolif 54:e12941 (2021). PubMed: 33111436
    • Brewer SM  et al. A Salmonella Typhi RNA thermosensor regulates virulence factors and innate immune evasion in response to host temperature. PLoS Pathog 17:e1009345 (2021). PubMed: 33651854
    • Li L  et al. Airborne particulate matter (PM2.5) triggers ocular hypertension and glaucoma through pyroptosis. Part Fibre Toxicol 18:10 (2021). PubMed: 33663554
    • Yue R  et al. NLRP3-mediated pyroptosis aggravates pressure overload-induced cardiac hypertrophy, fibrosis, and dysfunction in mice: cardioprotective role of irisin. Cell Death Discov 7:50 (2021). PubMed: 33723236
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-cleaved N-terminal GSDMD antibody [EPR20829-408]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (ls174t)
    Permeabilization
    Yes - 0.2%Triton
    Specification
    ls174t
    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-cleaved N-terminal GSDMD antibody [EPR20829-408]

    Excellent
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (GUT)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: edta
    Permeabilization
    No
    Specification
    GUT
    Blocking step
    BSA as blocking agent for 2 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

    Western blot abreview for Anti-cleaved N-terminal GSDMD antibody [EPR20829-408]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (differentiated THP-1 cells)
    Gel Running Conditions
    Reduced Denaturing (15%)
    Loading amount
    50 µg
    Treatment
    1 ug/ml staphylococcus aueus alpha toxin for 12 h
    Specification
    differentiated THP-1 cells
    Blocking step
    (agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    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 Jun 25 2021

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