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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)

Reviews (4) Submit a question References (40)
Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Immunoprecipitation - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
  • Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19859] to DFNA5/GSDME - N-terminal
  • Suitable for: WB, IP, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free PE

Overview

  • Product name

    Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal
    See all DFNA5/GSDME primary antibodies

  • Description

    Rabbit monoclonal [EPR19859] to DFNA5/GSDME - N-terminal

  • Host species

    Rabbit

  • Specificity

    Our previous testing data indicates that this antibody detects no signal or very weak signal in mouse heart, spleen, lung, stomach, testicle, skin, pancreas, muscle and lymph node tissues. Final results may be affected by the expression level of the tested samples.

  • Tested applications

    Suitable for: WB, IP, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse brain lysate; SH-SY5Y and EMT6 whole cell lysates; Human fetal brain lysate; Rat brain lysates; HEK-293 transfected with DDDDK tagged DFNA5/GSDME (N-terminal) expression vector whole cell lysate; SGC-7901 cell lysate. Flow Cyt (intra).: SH-SY5Y and EMT6 cells. IP: EMT6 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: 0.05% BSA, 40% Glycerol, PBS
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR19859

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab215191 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 (3)
1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).

Our previous testing data indicates that this antibody detects no signal or very weak signal in mouse heart, spleen, lung, stomach, testicle, skin, pancreas, muscle and lymph node tissues. Final results may be affected by the expression level of the tested samples.
IP
1/30.
Flow Cyt (Intra)
1/60.
Notes
WB
1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).

Our previous testing data indicates that this antibody detects no signal or very weak signal in mouse heart, spleen, lung, stomach, testicle, skin, pancreas, muscle and lymph node tissues. Final results may be affected by the expression level of the tested samples.
IP
1/30.
Flow Cyt (Intra)
1/60.

Target

  • Tissue specificity

    Expressed in cochlea. Low level of expression in heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas, with highest expression in placenta.

  • Involvement in disease

    Defects in DFNA5 are the cause of deafness autosomal dominant type 5 (DFNA5) [MIM:600994]. DFNA5 is a form of sensorineural hearing loss. Sensorineural deafness results from damage to the neural receptors of the inner ear, the nerve pathways to the brain, or the area of the brain that receives sound information.

  • Sequence similarities

    Belongs to the gasdermin family.
  • Information by UniProt

  • Database links

    • Alternative names

      • 2310037D07Rik antibody
      • 4932441K13Rik antibody
      • Deafness, autosomal dominant 5 antibody
      • Deafness, autosomal dominant 5 protein antibody
      • DFNA5 antibody
      • DFNA5 gene antibody
      • DFNA5_HUMAN antibody
      • Dfna5h antibody
      • EG14210 antibody
      • Fin15 antibody
      • ICERE 1 antibody
      • ICERE-1 antibody
      • Inversely correlated with estrogen receptor expression 1 antibody
      • Non-syndromic hearing impairment protein 5 antibody
      • Nonsyndromic hearing impairment protein antibody
      see all

    Images

    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/1000 dilution

      Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates with 5% NFDM/TBST
      Lane 2 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST

      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: 55 kDa
      Observed band size: 55 kDa



      Exposure times:

      Lane 1: 15s

      Lane 2: 3 min

      The band around 35kDa is GSDME-NT which might be cleaved by activated caspase-3 (PMID: 33589596, PMID: 31347748,PMID: 35623158).

    • Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized EMT6 (mouse mammary carcinoma cell line) cell line labeling DFNA5/GSDMEwith ab215191 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) (ab150077) at 1/2000 dilution was used as the secondary antibody. 

      EMT6 cells were kindly provided by our collaborator Dr. Feng Shao, NIBS 

       

       

    • Immunoprecipitation - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Immunoprecipitation - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)

      ab215191 at 1/30 immunoprecipitating DFNA5/GSDME in EMT6 (mouse mammary carcinoma cell line) whole cell lysate.

      Lane 1 (input): EMT6 whole cell lysate (10μg)

      Lane 2 (+): ab215191 + EMT6 whole cell lysate

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of  ab215191 in EMT6 whole cell lysate

      For western blotting, ab215191 at 1/500. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

      EMT6 cells were kindly provided by our collaborator Dr. Feng Shao, NIBS

    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 0.7 µg/ml

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Mouse kidney tissue lysate
      Lane 3 : Mouse liver tissue lysate
      Lane 4 : Mouse thymus tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG) at 1/2000 dilution

      Predicted band size: 55 kDa
      Observed band size: 55 kDa


      Exposure time: 3 minutes


      Blocking and diluting buffer: 5% NFDM/TBST

    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)Image from Wang Y et al., Biochem Biophys Res Commu. 2018;495(1):1418-1425. Fig 4(A).; 10.1016/j.bbrc.2017.11.156 with permission from Elsevier.
      All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/1000 dilution

      Lane 1 : Wild Type SGC-7901 (human papillomavirus-related endocervical adenocarcinoma cell line) cell lysates
      Lane 2 : DFNA5/GSDME Knockout SGC-7901 cell lysates

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution

      Predicted band size: 55 kDa



      https://www.sciencedirect.com/journal/biochemical-and-biophysical-research-communications

    • Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized SH-SY5Y (human neuroblastoma cell line from bone marrow) cell line labeling DFNA5/GSDME with ab215191 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Different batches of ab215191 were tested on SH-SY5Y (Human neuroblastoma epithelial cell) lysate at 0.6 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 55 kDa.
    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/5000 dilution

      Lane 1 : SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate
      Lane 2 : DFNA5/GSDME knockout SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate

      Lysates/proteins at 10 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: 55 kDa
      Observed band size: 55 kDa


      Exposure time: 15 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The cell lysates were kindly provided by our collaborator, Dr. Feng Shao, NIBS.

    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/10000 dilution

      Lane 1 : HEK-293 (human epithelial cell line from embryonic kidney) transfected with a control vector containing DDDDK tag, whole cell lysate
      Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) transfected with DDDDK tagged DFNA5/GSDME (N-terminal) expression vector, 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: 55 kDa
      Observed band size: 37 kDa why is the actual band size different from the predicted?


      Exposure time: 1 second


      Blocking/Dilution buffer: 5% NFDM/TBST.

      This antibody is specific to the N-terminus of DFNA5/GSDME.

      The cell lysates were kindly provided by our collaborator, Dr. Feng Shao, NIBS.

    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/1000 dilution + Human fetal brain lysate at 10 µg

      Secondary
      VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 55 kDa
      Observed band size: 55 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular weight observed is consistent with what has been described in the literature (PMID: 28459430).

    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/1000 dilution

      Lane 1 : EMT6 (mouse mammary carcinoma cell line) whole cell lysate
      Lane 2 : Mouse brain lysate
      Lane 3 : Rat brain lysate

      Lysates/proteins at 10 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: 55 kDa
      Observed band size: 55 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular weight observed is consistent with what has been described in the literature (PMID: 28459430).

      The EMT6 cell lysate was kindly provided by our collaborator Dr. Feng Shao, NIBS.

    • Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
      Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)

    References (40)

    Publishing research using ab215191? Please let us know so that we can cite the reference in this datasheet.

    ab215191 has been referenced in 40 publications.

    • Zhang Q  et al. Leptin induces NAFLD progression through infiltrated CD8+ T lymphocytes mediating pyroptotic-like cell death of hepatocytes and macrophages. Dig Liver Dis 53:598-605 (2021). PubMed: 33172809
    • Tang J  et al. Acute cadmium exposure induces GSDME-mediated pyroptosis in triple-negative breast cancer cells through ROS generation and NLRP3 inflammasome pathway activation. Environ Toxicol Pharmacol 87:103686 (2021). PubMed: 34098069
    • Zhang Y  et al. A unique death pathway keeps RIPK1 D325A mutant mice in check at embryonic day 10.5. PLoS Biol 19:e3001304 (2021). PubMed: 34437534
    • Chen YR  et al. siRNAs Targeting Mouse-Specific lncRNA AA388235 Induce Human Tumor Cell Pyroptosis/Apoptosis. Front Oncol 11:662444 (2021). PubMed: 34195074
    • Tamura Y  et al. Anti-pyroptotic function of TGF-ß is suppressed by a synthetic dsRNA analogue in triple negative breast cancer cells. Mol Oncol 15:1289-1307 (2021). PubMed: 33342034
    View all Publications for this product

    Customer reviews and Q&As

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    1-4 of 4 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal

     Average
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (GUT)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA
    Permeabilization
    No
    Specification
    GUT
    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

    Western blot abreview for Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Monkey Cell lysate - whole cell (monkey Retinal Pigmented Epithelium cells)
    Gel Running Conditions
    Reduced Denaturing (4-12 % NuPAGE® Novex®Bis-Tris Gels)
    Loading amount
    20 µg
    Specification
    monkey Retinal Pigmented Epithelium cells
    Blocking step
    Milk as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°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 Dec 01 2020

    Western blot abreview for Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal

     Excellent
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Lysates of BMDCs)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    25000 cells
    Treatment
    LPS (3h, 50 ng/ml), +/- Nigericin
    Specification
    Lysates of BMDCs
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°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 Aug 23 2020

    Western blot abreview for Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (EMT6)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    EMT6
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°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 Sep 26 2017

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