Analysis Note

ControlCardiac tissue, particularly sinoatrial tissues.

Application

Anti-Connexin 45 Antibody, near C-terminus, cytoplasmic, clone 8A11.2 is an antibody against Connexin 45 for use in IC, IH & WB.

Immunblotting (~46 kDa): 0.5 -1 µg/mL with ECL detection. Samples should be prepared with proteinase inhibitors and kept cold for best results. Alkali extraction methods can be used to enrich for connexin protein fractions. Briefly, 1 ml of NaHCO3 containing protease and phosphatase inhibitors and 22 µl of 1 M NaOH were added to frozen cell pellets. This cell suspension was sonicated for 30 s and incubated on ice for 50 min. Then, the homogenate was centrifuged for 30 min at 30,000 X g at 4°C. The supernatant was discarded, and the pellet was resuspended in sample buffer. {Elenes, S et al 2001}. 8% SDS_PAGE gels and PVDF membranes are suggested for greatest clarity.

Immunohistochemistry: fresh frozen, methanol fixed sections {Ujiie H et al (2003)}; 4% fixed frozen sections {Ikeda, Y et al 2007). 1-10 µg/mL with overnight incubations; 4% PFA fixed, paraffin-embedded sections {Gluhak-Heinrich, J et al 2007}, 1:50 with enzymatic detection.

Immunocytochemistry: 2% PFA fixed cells (15', RT), permeabilized with a solution of 0.2% triton X-100/4% BSA {Sorensen et al 2008}.


Optimal working dilutions must be determined by end user.

Research CategoryCell Structure

Research Sub CategoryAdhesion (CAMs)

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Connexin 45 is member of a family of gap junction proteins known as Connexin gene family. It is expressed in a wide variety of cell types during development and adult life, particularly in cardiac and smooth muscle tissues as well as brain tissues. Connexin 45 forms part of a network of intercellular channels that provide a route for the diffusion of low molecular weight materials from cell to cell. There a number of alternatively spliced transcript variants that all seen to encode the same protein isoform.

Immunogen

The peptide sequence used for the immunogen was H-Gln-Ala-Tyr-Ser-His-Gln-Asn-Asn-Pro-His-Gly-Pro-Arg-Glu-Cys-OH conjugated to KLH, corresponding to amino acids 354-367 of human connexin 45.

Epitope: near C-terminus, cytoplasmic

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Format: Purified

Liquid at 1 mg/mL in 0.02M PB with 0.25M NaCl,pH=7.6 containing 0.1% sodium azide as a preservative.

Specificity

Others species not tested. Also confirmed to work on rat C6 cells.

Clone 8A11.2 is highly specific for Connexin 45 protein with no cross reactivity as analyzed by western blotting to any other connexin family member.

Storage and Stability

Maintain at 2° to 8°C in undiluted aliquots for up to 12 months fromd date of receipt. Prevent multiple warming and coolings. Aliquot in tightly closed tubes to prevent loss.