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使用敲除细胞株进行验证重组RabMAb

重组Anti-DFNA5/GSDME抗体[EPR19859] - BSA and Azide free (ab223877)

Submit a review Submit a question References (1)
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)
  • 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

概述

  • 产品名称

    Anti-DFNA5/GSDME抗体[EPR19859] - BSA and Azide free
    参阅全部 DFNA5/GSDME 一抗

  • 描述

    兔单克隆抗体[EPR19859] to DFNA5/GSDME - BSA and Azide free

  • 宿主

    Rabbit

  • 特异性

    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.

  • 经测试应用

    适用于: IP, Flow Cyt (Intra), WBmore details

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

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

  • 阳性对照

    • 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.

  • 常规说明

    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.

性能

  • 形式

    Liquid

  • 存放说明

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • 存储溶液

    pH: 7.2
    Constituent: PBS

  • 无载体

    是
  • Concentration information loading...

  • 纯度

    Protein A purified

  • 克隆

    单克隆

  • 克隆编号

    EPR19859

  • 同种型

    IgG

  • 研究领域

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab223877于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
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).
说明
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).

靶标

  • 组织特异性

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

  • 疾病相关

    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.

  • 序列相似性

    Belongs to the gasdermin family.
  • Information by UniProt

  • 数据库链接

    • 别名

      • 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

    图片

    • Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
      Western blot - Anti-DFNA5/GSDME antibody [EPR19859] - BSA and Azide free (ab223877)
      Lanes 1 & 4 : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/1000 dilution
      Lanes 2 & 5 : Anti-DFNA5/GSDME antibody [EPR19859-60] - C-terminal (ab221843) at 1/1000 dilution
      Lanes 3 & 6 : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/1000000 dilution

      Lanes 1-3 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
      Lanes 4-6 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate

      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



      Blocking and dilution buffer: 5% NFDM/TBST.

    • 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)

    文献 (1)

    发表研究结果有使用 ab223877？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab223877 被引用在 1 文献中.

    • 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

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