Astana Biomed Group, an authorized Abcam distributor in Central Asia

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, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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 & Species

    Application Species
    Flow Cyt
    Mouse
    Human
    IP
    Mouse
    WB
    Mouse
    Human
    See all applications and species data

  • 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.: 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

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

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

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized EMT6 (mouse mammary carcinoma cell line) cell line labeling DFNA5/GSDME with 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 - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
    Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)

    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)
    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)

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