Recombinant
RabMAb

Recombinant Anti-GSDMA antibody [EPR19858-104] (ab230768)

Overview

  • Product name

    Anti-GSDMA antibody [EPR19858-104]
    See all GSDMA primary antibodies
  • Description

    Rabbit monoclonal [EPR19858-104] to GSDMA
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein within Human GSDMA aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q96QA5

  • Positive control

    • WB: Hela cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate; Hela cells stably transfected with mouse GSDMA expression vector, containing a DDDDK-tag, whole cell lysate; mouse skin and stomach tissue lysate; human skin and stomach tissue lysate; rat skin tissue lysate. IP: Hela cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate. IHC-P: Human skin tissue; mouse stomach tissue; rat stomach tissue.
  • 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: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19858-104
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab230768 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
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 49 kDa).
IP 1/30.

Target

  • Relevance

    Function: May promote pyroptosis (Probable). Upon cleavage in vitro of genetically engineered GSDMA, the released N-terminal moiety binds to some types of lipids, such as possibly phosphatidylinositol (4,5)-bisphosphate. Homooligomerizes within the membrane and forms pores of 10 -15 nanometers (nm) of inner diameter, triggering cell death. Also binds to bacterial and mitochondrial lipids, including cardiolipin, and exhibits bactericidal activity (PubMed:27281216). The physiological relevance of these observations is unknown. Tissue specificity: Expressed predominantly in the gastrointestinal tract and, at a lower level, in the skin. Also detected in mammary gland. In the gastrointestinal tract, mainly expressed in differentiated cells, including the differentiated cell layer of esophagus and mucus-secreting pit cells of the gastric epithelium. Down-regulateded in gastric cancer cells. Similarity: Belongs to the gasdermin family. Domain: Intramolecular interactions between N- and C-terminal domains may be important for autoinhibition in the absence of activation signal. The intrinsic pyroptosis-inducing activity is carried by the N-terminal domain.
  • Cellular localization

    Cytoplasm, perinuclear region
  • Database links

  • Alternative names

    • FKSG9 antibody
    • Gasdermin 1 antibody
    • Gasdermin A antibody
    • Gasdermin-1 antibody
    • Gasdermin-A antibody
    • GSDM antibody
    • GSDM1 antibody
    • GSDMA antibody
    • GSDMA_HUMAN antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling GSDMA with ab230768 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on squamous epithelium of rat stomach (PMID: 23979942) 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.

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-GSDMA antibody [EPR19858-104] (ab230768) at 1/1000 dilution

    Lane 1 : Mouse skin tissue lysate
    Lane 2 : Mouse stomach tissue lysate
    Lane 3 : Human skin tissue lysate
    Lane 4 : Human stomach tissue lysate
    Lane 5 : Rat skin tissue lysate

    Lysates/proteins at 20 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 49 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times.

    Lane 1: 32 seconds.

    Lanes 2 & 3: 3 minutes.

    Lanes 4 & 5: 102 seconds.

    This image is produced using super sensitivity ECL substrate. We strongly suggest the customer to use higher sensitivity ECL substrate when developing the blot.

  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling GSDMA with ab230768 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on squamous epithelium of mouse stomach (PMID: 23979942) 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.

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human skin tissue labeling GSDMA with ab230768 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on squamous epithelium of human skin (PMID: 23979942) 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.

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • GSDMA was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate with ab230768 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230768 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Lane 1: Hela cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate 10 μg (Input).
    Lane 2: ab230768 IP in Hela cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate. 
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab230768 in Hela cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: less than 1 second.

    The observed molecular mass is consistent with the literature (PMID:17350798).

    The observed molecular masses lower than 50 kDa are degraded expressed GSDMA protein. The cells were kindly provided by our collaborator Dr. Feng Shao, NIBS.

  • All lanes : Anti-GSDMA antibody [EPR19858-104] (ab230768) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Hela cells stably transfected with mouse GSDMA2 expression vector, containing a DDDDK-tag, whole cell lysate
    Lane 3 : Hela cells stably transfected with mouse GSDMA3 expression vector, containing a DDDDK-tag, 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

    Developed using the ECL technique.

    Predicted band size: 49 kDa


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-GSDMA antibody [EPR19858-104] (ab230768) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Hela cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate
    Lane 3 : Hela cells stably transfected with mouse GSDMA expression vector, containing a DDDDK-tag, 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

    Developed using the ECL technique.

    Predicted band size: 49 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

    The observed molecular mass is consistent with the literature (PMID:17350798).
    The observed molecular masses lower than 50 kDa are degraded expressed GSDMA protein. The cells were kindly provided by our collaborator Dr. Feng Shao, NIBS.

References

ab230768 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Hela GSDMA-KO or Stable)
Total protein in input
5000 µg
Immuno-precipitation step
Protein A
Specification
Hela GSDMA-KO or Stable

Abcam user community

Verified customer

Submitted Jul 01 2019

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

Sign up