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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)

Reviews (1) Submit a question References (4)
Western blot - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
  • Western blot - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
  • Immunoprecipitation - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
  • Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21153] to Connexin 43 / GJA1
  • Suitable for: IHC-P, IP, WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Connexin 43 / GJA1 antibody [EPR21153]
    See all Connexin 43 / GJA1 primary antibodies

  • Description

    Rabbit monoclonal [EPR21153] to Connexin 43 / GJA1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, IP, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa whole cell lysate; Human testis, fetal brain and breast cancer lysates. IP: Human testis tissue lysate. IHC-P: Human cardiac muscle and prostate tissue.

  • 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab217676 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
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
WB (1)
1/1000. Predicted molecular weight: 43 kDa.
Notes
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
WB
1/1000. Predicted molecular weight: 43 kDa.

Target

  • Function

    One gap junction consists of a cluster of closely packed pairs of transmembrane channels, the connexons, through which materials of low MW diffuse from one cell to a neighboring cell. May play a critical role in the physiology of hearing by participating in the recycling of potassium to the cochlear endolymph.

  • Tissue specificity

    Expressed in the heart and fetal cochlea.

  • Involvement in disease

    Defects in GJA1 are the cause of autosomal dominant oculodentodigital dysplasia (ODDD) [MIM:164200]; also known as oculodentoosseous dysplasia. ODDD is a highly penetrant syndrome presenting with craniofacial (ocular, nasal, dental) and limb dysmorphisms, spastic paraplegia, and neurodegeneration. Craniofacial anomalies tipically include a thin nose with hypoplastic alae nasi, small anteverted nares, prominent columnella, and microcephaly. Brittle nails and hair abnormalities of hypotrichosis and slow growth are present. Ocular defects include microphthalmia, microcornea, cataracts, glaucoma, and optic atrophy. Syndactyly type 3 and conductive deafness can occur in some cases. Cardiac abnormalities are observed in rare instances.
    Defects in GJA1 are the cause of autosomal recessive oculodentodigital dysplasia (ODDD autosomal recessive) [MIM:257850].
    Defects in GJA1 may be the cause of syndactyly type 3 (SDTY3) [MIM:186100]. Syndactyly is an autosomal dominant trait and is the most common congenital anomaly of the hand or foot. It is marked by persistence of the webbing between adjacent digits, so they are more or less completely attached. In this type there is usually complete and bilateral syndactyly between the fourth and fifth fingers. Usually it is soft tissue syndactyly but occasionally the distal phalanges are fused. The fifth finger is short with absent or rudimentary middle phalanx. The feet are not affected.
    Defects in GJA1 are a cause of hypoplastic left heart syndrome (HLHS) [MIM:241550]. HLHS refers to the abnormal development of the left-sided cardiac structures, resulting in obstruction to blood flow from the left ventricular outflow tract. In addition, the syndrome includes underdevelopment of the left ventricle, aorta, and aortic arch, as well as mitral atresia or stenosis.
    Defects in GJA1 are a cause of Hallermann-Streiff syndrome (HSS) [MIM:234100]. HSS is a disorder characterized by a typical skull shape (brachycephaly with frontal bossing), hypotrichosis, microphthalmia, cataracts, beaked nose, micrognathia, skin atrophy, dental anomalies and proportionate short stature. Mental retardation is present in a minority of cases.

  • Sequence similarities

    Belongs to the connexin family. Alpha-type (group II) subfamily.

  • Cellular localization

    Cell membrane. Cell junction > gap junction.
  • Information by UniProt

  • Database links

    • Alternative names

      • Connexin 43 antibody
      • Connexin-43 antibody
      • Cx 43 antibody
      • Cx43 antibody
      • CXA1_HUMAN antibody
      • DFNB38 antibody
      • Gap junction 43 kDa heart protein antibody
      • Gap junction alpha-1 protein antibody
      • Gap junction protein alpha 1 43kDa (connexin 43) antibody
      • Gap junction protein alpha 1 43kDa antibody
      • Gap junction protein alpha like antibody
      • GJA 1 antibody
      • Gja1 antibody
      • GJAL antibody
      • ODD antibody
      • ODDD antibody
      • ODOD antibody
      • SDTY3 antibody
      see all

    Images

    • Western blot - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
      Western blot - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
      All lanes : Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
      Lane 2 : GJA1 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
      Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 43 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab217676 observed at 43 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab217676 was shown to recognize Connexin 43 / GJA1 in wild-type HEK 293 cells as signal was lost at the expected MW in GJA1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GJA1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab217676 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
      Western blot - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
      All lanes : Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676) at 1/1000 dilution

      Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : Human testis tissue lysate
      Lane 3 : Human fetal brain tissue lysate
      Lane 4 : Human breast cancer tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 43 kDa
      Observed band size: 26-32 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times.
      Lane 1: 37 seconds.
      Lane 2: 15 seconds.
      Lane 3: 103 seconds.
      Lane 4: 3 minutes.

      The expression profile is consistent with the existence of natural variants described in the literature (PMID: 24210816; PMID: 28576298).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)

      Immunohistochemical analysis of paraffin-embedded human prostate tissue labeling Connexin 43 / GJA1 with ab217676 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use. Positive staining on basal cells of human prostate gland (PMID: 20735413) is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)

      Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue labeling Connexin 43 / GJA1 with ab217676 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use. Positive staining on human cardiac muscle (PMID: 25018732) is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunoprecipitation - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
      Immunoprecipitation - Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)

      Connexin 43 / GJA1 was immunoprecipitated from 0.35 mg of human testis lysate with ab217676 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217676 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: Human testis lysate 10 μg (Input).
      Lane 2: ab217676 IP in human testis lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab217676 in human testis lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 1 minute.

      The expression profile is consistent with the existence of natural variants described in the literature (PMID: 24210816; PMID: 28576298).

    • Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)
      Anti-Connexin 43 / GJA1 antibody [EPR21153] (ab217676)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (4)

    Publishing research using ab217676? Please let us know so that we can cite the reference in this datasheet.

    ab217676 has been referenced in 4 publications.

    • Fan J  et al. lncRNA LEF1-AS1 Acts as a Novel Biomarker and Promotes Hypopharyngeal Squamous Cell Carcinoma Progression and Metastasis by Targeting the miR-221-5p/GJA1 Axis. Dis Markers 2022:3881310 (2022). PubMed: 35371339
    • Deng W  et al. Proteomic and miRNA Profiles of Exosomes Derived from Myometrial Tissue in Laboring Women. Int J Mol Sci 23:N/A (2022). PubMed: 36293200
    • Wang L  et al. Triiodothyronine and dexamethasone alter potassium channel expression and promote electrophysiological maturation of human-induced pluripotent stem cell-derived cardiomyocytes. J Mol Cell Cardiol 161:130-138 (2021). PubMed: 34400182
    • Blume GG  et al. Bone-Marrow Stem Cells and Acellular Human Amniotic Membrane in a Rat Model of Heart Failure. Life (Basel) 11:N/A (2021). PubMed: 34575107

    Customer reviews and Q&As

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    Western blot abreview for Anti-Connexin 43 / GJA1 antibody [EPR21153]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Lung)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    20 µg
    Specification
    Lung
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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    Verified customer

    Submitted Mar 22 2022

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