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  1. Link

    dfna5gsdme-antibody-epr19859-bsa-and-azide-free-ab223877.pdf

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Validated using a knockout cell lineRecombinantRabMAb

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

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  • Certificate of Compliance
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Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Immunoprecipitation - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
  • Flow Cytometry (Intracellular) - 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 (Intra), WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 PE Unconjugated

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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 (Intra), WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • 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 (intra).: SH-SY5Y cells.
  • General notes

    ab223877 is the carrier-free version of ab215191.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    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.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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.

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

The Abpromise guarantee

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 (Intra)
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).
Notes
IP
Use at an assay dependent concentration.
Flow Cyt (Intra)
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
    • 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

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

      Intracellular 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). 

       

       

    • 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).

    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
      This data was developed using ab215191, the same antibody clone in a different buffer formulation. 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.
    • Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
      Flow Cytometry (Intracellular) - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)

      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. 

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

       

       

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

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized SH-SY5Y (human neuroblastoma cell line from bone marrow) cell line labeling DFNA5/GSDMEwith 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 download

      Download

    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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