Validated using a knockout cell line
Recombinant
RabMAb

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

Knockout Tested Rabbit recombinant monoclonal DFNA5/GSDME antibody [EPR19859]. Validated in WB, IP, Flow Cyt and tested in Mouse, Rat, Human. Cited in 2 publication(s).

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, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human DFNA5/GSDME aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: O60443

  • Positive control
    • WB: Mouse brain, and kidney lysates; 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

    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: 0.05% BSA, 40% Glycerol, PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR19859
  • Isotype
    IgG
  • Research areas

Applications

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 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.

Flow Cyt 1/60.
IP 1/30.

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

  • 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 cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized EMT6 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

  • 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

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

    Lane 1 : Wild Type SGC-7901 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 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.

  • 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.

  • ab215191 at 1/30 immunoprecipitating DFNA5/GSDME in EMT6 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

  • 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.

  • 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).

  • 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.

References

This product has been referenced in:
  • Zhang CC  et al. Chemotherapeutic paclitaxel and cisplatin differentially induce pyroptosis in A549 lung cancer cells via caspase-3/GSDME activation. Apoptosis N/A:N/A (2019). Read more (PubMed: 30710195) »
  • Wu M  et al. A PLK1 kinase inhibitor enhances the chemosensitivity of cisplatin by inducing pyroptosis in oesophageal squamous cell carcinoma. EBioMedicine 41:244-255 (2019). Read more (PubMed: 30876762) »
See all 6 Publications for this product

Customer reviews and Q&As

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

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Verified customer

Submitted Sep 26 2017

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