Monoclonal Anti-Calbindin-D-28K antibody produced in mouse

Calbindin-D-28K is a highly conserved 28 kDa calcium binding protein with broad tissue distribution.

Monoclonal Anti-Calbindin-D-28K (mouse IgG1 isotype) is derived from the CB-955 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from BALB/c mice immunized with a purified bovine kidney calbindin-D-28K. The isotype is determined by a double diffusion immunoassay using Mouse Monoclonal Antibody Isotyping Reagents, Catalog Number ISO2.

Calbindin-D-28K belongs, together with calmodulin, S-100, parvalbumin, troponin C and other proteins, to a family of low molecular weight calcium-binding proteins (CaBPs). These CaBPs have homologous primary structures, which contain polypeptide folds of the EF-hand type for the acceptance of Ca²⁺. Calbindin is an attractive neuroanatomical marker. Despite its broad tissue distribution, calbindin-D-28K exhibits a cell-type specific expression pattern. Thus, it has been immunocytochemically localized in selected cells in many CNS structures, where it is thought to act either as an intracellular calcium buffer, or in the intramembranous transport of calcium. It 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. Monoclonal antibody reacting specifically with calbindin-D-28K is instrumental in the localization of calbindin in different tissues and cell populations.

The antibody does not react with other members of the EF-hand family, such as calbindin-D-9K, calretinin, myosin light chain, parvalbumin, S-100a, S-100b, S-100A2 (S100L) and S-100A6 (calcyclin). A weaker reactivity was observed with chicken calbindin-D-28K.

Bovine kidney calbindin-D

Monoclonal Anti-Calbindin-D-28K antibody produced in mouse may be used in immunohistochemistry (alcohol-, formalin-, and methacarn-fixed, paraffin-embedded, and frozen tissue sections). Enzymatic predigestion of formalin-fixed, paraffin-embedded sections improves immunohistochemical staining. The antibody produced in mouse is suitable for indirect ELISA, western blotting at a working dilution of 1:3000 using MDBK cell extract and indirect immunoperoxidase staining with a working dilution of 1:3000 using trypsin-digested, formalin-fixed, paraffin-embedded rat cerebellum sections. It was used for immunohistochemistry and immunocytochemistry at a working dilution of 1:500 in a study to understand impaired cerebellar development and function in mice lacking CAPS2, a protein involved in neurotrophin release. It was used for immunostaining to gauge the development of neuronal and glial cells in the inner nuclear layer of islet-class factor Islet-1 (Isl1)-null and control retinas. It was used for immunohistochemical colocalization studies to confirm postsynaptic localization of the RGS11 complex in a study.

Calbindin-D-28K is involved in intramembranous transport of calcium or acts as an intracellualr calcium buffer in many CNS structures such as subpopulations of central and peripheral nervous system neurons. It is also present 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.

Reagent is supplied as ascites fluid with 15mM sodium azide as a preservative.

For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing, or storage in "frost free" freezers, is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.

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