Recombinant
RabMAb

Recombinant Anti-Connexin 43 / GJA1 antibody [EPR22955-101] (ab235585)

Overview

  • Product name

    Anti-Connexin 43 / GJA1 antibody [EPR22955-101]
    See all Connexin 43 / GJA1 primary antibodies
  • Description

    Rabbit monoclonal [EPR22955-101] to Connexin 43 / GJA1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, IHC-Fr, IHC-P, WBmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Connexin 43/ GJA1 aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: P17302

  • Positive control

    • WB: Mouse brain, mouse heart, rat brain and human brain lysates. IHC-P: Human, mouse and rat cardiac muscle tissues. Mouse testis tissue. IP: Mouse brain 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

Applications

Our Abpromise guarantee covers the use of ab235585 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 1/30.
IHC-Fr 1/250.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Use at 1/500 for human.

Use at 1/1000 for mouse and rat.

WB 1/1000.
  • Application notes
    Is unsuitable for Flow Cyt or ICC/IF.
  • 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

    • All lanes : Anti-Connexin 43 / GJA1 antibody [EPR22955-101] (ab235585) at 1/1000 dilution

      Lane 1 : Mouse brain tissue lysate at 20 µg
      Lane 2 : Mouse heart tissue lysate at 20 µg
      Lane 3 : Rat brain tissue lysate at 10 µg
      Lane 4 : Human brain tissue lysate at 10 µg

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

      Observed band size: 43 kDa
      why is the actual band size different from the predicted?



      Blocking and Dilution Buffer and concentration: 5% NFDM/TBST. The expression profile is consistent with the existence of natural variants described in the literature (PMID: 24210816, 28576298). Exposure times: Lane 1: 59 seconds; Lane 2: 15 seconds; Lane 3: 15 seconds; Lane 4: 26 seconds.

    • Connexin 43 / GJA1 was immunoprecipitated from 0.35 mg mouse brain lysate 10µg with ab235585 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab235585 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: Mouse brain lysate 10µg.

      Lane 2: ab235585 IP in mouse brain lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab235585 in mouse brain lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 10 seconds.

      High MW band between 75kDa and 100kDa represents Connexin 43 dimers which described in literature. (PMID: 19460776).

       

    • Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue labeling Connexin 43 / GJA1 with ab235585 at 1/1000 dilution (0.507 µg/ml) followed by a ready to use Goat Anti-Rabbit & Mouse IgG (HRP). Positive staining on rat myocardial intercalated disc is observed. The section was incubated with ab235585 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit & Mouse IgG (HRP).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Connexin 43 / GJA1 with ab235585 at 1/1000 dilution (0.507 µg/ml) followed by a ready to use Goat Anti-Rabbit & Mouse IgG (HRP). Membranous staining on mouse testis is observed. The section was incubated with ab235585 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit & Mouse IgG (HRP).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue labeling Connexin 43 / GJA1 with ab235585 at 1/1000 dilution (0.507 µg/ml) followed by a ready to use Goat Anti-Rabbit & Mouse IgG (HRP). Positive staining on mouse myocardial intercalated disc (PMID: 30228305) is observed. The section was incubated with ab235585 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit & Mouse IgG (HRP).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue labeling Connexin 43 / GJA1 with ab235585 at 1/500 dilution (1.01 µg/ml) followed by a ready to use Goat Anti-Rabbit & Mouse IgG (HRP). Positive staining on human myocardial intercalated disc is observed. The section was incubated with ab235585 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit & Mouse IgG (HRP).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen section of rat cardiac muscle tissue labeling Connexin 43 / GJA1 using ab235585 at 1/250 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). The nuclear counterstain is DAPI (blue). Positive staining on intercalated discs of rat cardiac muscle (PMID: 18678643) is observed.

      Secondary antibody only control: Used PBS instead of primary antibody, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

       

    • 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen section of mouse cardiac muscle tissue labeling Connexin 43 / GJA1 using ab235585 at 1/250 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). The nuclear counterstain is DAPI (blue). Positive staining on intercalated discs of mouse cardiac muscle (PMID: 18678643) is observed.

      Secondary antibody only control: Used PBS instead of primary antibody, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

    References

    ab235585 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    There are currently no Customer reviews or Questions for ab235585.
    Please use the links above to contact us or submit feedback about this product.

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Sign up