Cx43 antibody | knockout validation | Sigma-Aldrich C6219
DOI
//dx.doi.org/10.13070/ko.en.6.1742
Date
2016-10-18

This is a knockout-validated antibody summary, based on the publication "New protein–protein interactions of mitochondrial connexin 43 in mouse heart", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Cx43 antibody | knockout validation | Sigma-Aldrich C6219 figure 1
Figure 1. Western Blot analysis of Cx43 in WT and knock-in Cx43KICx32 (KI) mice in total heart (THEART), subsarcolemmal (SSM) and interfibrillar (IFM) protein extracts. Anti‐SDHA was used as a mitochondrial loading control. From [1].
Antibody information

Rabbit polyclonal

Company: Sigma-Aldrich

Antibody: Cx43

Catalog number: C6219

Summary: Rabbit polyclonal raised against synthetic peptide corresponding to the C-terminal segment of the cytoplasmic domain (amino acids 363-382 with N-terminally added lysine) of human/rat connexin-43. Reacts with human and other mammals. Suitable for western blot, microarray, immunocytochemistry/immunofluorescence, immunohistochemistry (frozen and paraffin).

Cx43 antibody | knockout validation | Sigma-Aldrich C6219 figure 2
Figure 2. Confocal microscopy images of Cx43 (in red) showing a specific localization in cardiomyocyte membrane and mitochondria (SDHA protein, in green). Signal is completely absent in Cx43KICx32 cells. Nuclei is stained with Hoeschst 33342 (in blue). From [1].
Validation Method

Western blot | Immunocytochemistry

Sample

WB: WT and knock-in Cx43KICx32 (where the coding region of the Cx43 gene has been replaced by that of Cx32) mouse heart tissue. Total heart homogenates were attained by solubilization of 100–200 mg of tissue in lysis buffer [2].

IC: Adult cardiomyocytes isolated from WT and knock‐in Cx43KICx32 mice .

Blocking agent

IC: 1% BSA in PBS for 45 min at RT.

Secondary incubation

IC: AlexaFluor 594‐conjugated anti‐rabbit (A‐11012; Molecular Probes).

Detection

IC: Olympus Spectral Confocal Microscopy FV1000, Olympus, Center Valley, PA, USA.

Disclaimer

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.

References
  1. Denuc A, Núñez E, Calvo E, Loureiro M, Miro-Casas E, Guarás A, et al. New protein-protein interactions of mitochondrial connexin 43 in mouse heart. J Cell Mol Med. 2016;20:794-803 pubmed publisher
  2. 2% n‐Dodecyl β‐d‐maltoside (D4641; Sigma‐Aldrich), 20 mmol/L TEA pH 8, 20 mmol/L ethylenediaminetetraacetic acid, 10 mmol/L DTT and protease inhibitors cocktail.