Monoclonal anti-Na+-Ca2+-exchanger

 western blot, immunohistochemistry, immunocytochemistry, immunohistochemistry - frozen section, western blot knockout validation

 Mouse monoclonal, IgG1

 Conc. Supernat lyophil.

 Lyophilized, concentrated supernatant.

 with 200 µl of bidistilled water.

The monoclonal antibody against the canine Na+ -Ca2+-exchanger, recognizes two neighboring but non-overlapping sequences of the hydrophilic region of the exchanger, connecting the putative transmembrane segments 5 and 6. The antibody was evaluated for specificity and potency by immunoblotting, and immunoprecipitation (see reference). It recognizes virtually only NCX1 in monkey, horse, dog, rat, bovine and mouse tissue. At higher concentrations it reacts faintly with NCX2, whereas NCX3 is not recognized. Both splice variants with exon A (heart, nervous system) and exon B (kidney, glia) are recognized.

Immunohistochemistry: 1: 500; 

1: 1’000 Immunoblots and immunoprecipitation: 1: 500; 1: 1'000 (use milk for blocking). A strong 70 kDa band is found in addition to the 116 kDa of the integral exchanger. Often the HMW band is a double band 116/ 112 kDa band. 

The titer was determined using the indirect-peroxidase method. We recommend that the optimal dilutions be determined by titration experiments.

After reconstitution freeze in small aliquots (e.g. 1 µl) and keep at - 80°C (or at least - 20°C). For continuous use keep at 4°C (with 0.01% Na-azide). Avoid repeated freezing and thawing.

 Porzig H., Li Z., Nicoll D.A., Philipson K.D. (1993) Mapping the cardiac sodium-calcium exchanger with monoclonal antibodies. Am. J. Physiol. 265:C78 -C756, 1993