Calbindin, Recombinant, Rat (D28K, Vitamin D-dependent Calcium-binding Protein) discontinued

Molecular Biology / MB-Neuroscience

-20°C

Supplied as a lyophilized powder from 50mM sodium bicarbonate. Reconstitute with sterile dH2O to a concentration of ≥0.1mg/ml.

Calbindins are Ca-binding proteins belonging to the troponin C superfamily. CALB28K/Calbindin1/CALB1 (D28K/Spot35 protein or cholecalcin, rat 261 aa; mouse 261 aa; human 261-aa, chromosome 8q21.3-q22.1) was originally described as 27kD induced by vitamin D in the duodenum of chicken. In mammals, it is expressed in the kidney, pancreatic islets, and brain. In brain, its synthesis is independent of vitamin D. CABP28K contains 4 active and 2 inactive EF-hand Ca-binding domains. The gene for CABP28K is clustered in the same region as carbonic anhydrase. The neurons in the brains of patients with Huntington disease are CAB28K depleted. There are two types of CaBPs: the "trigger"- and the "buffer"-CaBPs. The conformation of "trigger" type CaBPs changes upon Ca2+ binding and exposes regions on protein that interact with target molecules, thus altering their activity. The buffer-type CABP are thought to control the intracellular calcium concentration. Calbindin D-28K is found predominantly in subpopulations of central and peripheral nervous system neurons, and in certain epithelial cells involved in Ca2+ transport such as distal tubular cells and cortical collecting tubules of the kidney, and in enteric neuroendocrine cells.

Applications:
CABP28K can be used for immunoblots, absorption experiments in immunohistochemistry, radioimmunoassay and intracellular injection. For adsorption we suggest the following procedure: A- Dilute 1 µl of the monoclonal antibody 300 against calbindin D-28k in 5 ml of the usual buffer for immunohistochemistry (final dilution 1:5000). B- Add 1ug of the recombinant protein to 1 ml of the diluted antibody solution and mix well. C- Incubate for at least 6 hours in the cold. D- Apply to tissue-sections and incubate for 3 days. E- Complete the immunohistochemical reaction as usual (biotinylated second antibody, ABC-complex, DAB). As a result, the immunostaining should be strongly reduced or even completely prevented.

Storage and Stability:
Lyophilized and reconstituted products are stable for 6 months after receipt at -20°C. Reconstitute with sterile ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.