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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Immunoprecipitation - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)

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 - BSA and Azide free
  • Suitable for: IP, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free
    See all DFNA5/GSDME primary antibodies
  • Description

    Rabbit monoclonal [EPR19859] to DFNA5/GSDME - BSA and Azide free
  • Host species

    Rabbit
  • Specificity

    Expression levels of the target protein vary with sample type and some optimisation may be required.

  • Tested applications

    Suitable for: IP, Flow Cyt, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: O60443

  • Positive control

    • WB: SH-SY5Y and EMT6 whole cell lysates; Human fetal brain lysate; Mouse and rat brain lysates; HEK-293 transfected with DDDDK tagged DFNA5/GSDME (N-terminal) expression vector whole cell lysate. Flow Cyt.: SH-SY5Y cells.
  • General notes

    Ab223877 is the carrier-free version of ab215191. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab223877 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19859
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Sensory System
    • Auditory system

Associated products

  • Alternative Versions

    • Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191)
    • Alexa Fluor® 488 Anti-DFNA5/GSDME antibody [EPR19859] (ab225519)
    • PE Anti-DFNA5/GSDME antibody [EPR19859] (ab225520)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human DFNA5/GSDME protein (ab116824)

Applications

Our Abpromise guarantee covers the use of ab223877 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
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).

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.
  • Target information above from: UniProt accession O60443 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1687 Human
    • Entrez Gene: 54722 Mouse
    • Entrez Gene: 353316 Rat
    • Omim: 608798 Human
    • SwissProt: O60443 Human
    • SwissProt: Q9Z2D3 Mouse
    • Unigene: 520708 Human
    • Unigene: 248361 Mouse
    • Unigene: 96433 Rat
    see all
  • 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] - BSA and Azide free (ab223877)
    Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)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 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



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215191).

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

  • Immunoprecipitation - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
    Immunoprecipitation - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)

    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 and ab131366 VeriBlot for IP Detection Reagent (HRP) was used at 1/1000.

    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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215191).

  • Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
    Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)

    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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215191).

  • Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
    Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215191).

  • Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
    Flow Cytometry - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)

    This Flow Cyt data was generated using the same anti-DFNA5/GSDME antibody clone [EPR19859] in a different buffer formulation (cat# 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.

  • Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
    Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)

Protocols

  • Western blot protocols
  • Flow cytometry protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet

    Certificate of Compliance

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

  • References (1)

    Publishing research using ab223877? Please let us know so that we can cite the reference in this datasheet.

    ab223877 has been referenced in 1 publication.

    • Wu YL  et al. Microcystin-LR promotes necroptosis in primary mouse hepatocytes by overproducing reactive oxygen species. Toxicol Appl Pharmacol 377:114626 (2019). PubMed: 31201821

    Customer reviews and Q&As

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