Calretinin, mouse monoclonal, Cat# MCA-6A9

Calretinin, mouse monoclonal, Cat# MCA-6A9


Left: Mixed neuron/glial cultures stained with MCA-6A9 at 1:2,000 in red, and our chicken polyclonal antibody to Vimentin (CPCA-Vim) in green. Calretinin is prominently expressed in small number of interneurons, while astrocytes and fibroblasts were visualized with the vimentin antibody. Middle: Adult rat cortical section (45 μM; fixed by transcardial perfusion with 4% paraformaldehyde) was co-stained with MCA-6A9 (1:1000; red) and our chicken polyclonal antibody to calbinidin (CPCA-Calbindin; green). In the motor cortex, calretinin is expressed in a small population of interneurons that do not express calbindin. Because each antibody specifically labels a different population of cells exclusively, the cells are either stained with red, or green. Right: Adult mouse brain hippocampal section (45 μM; fixed by transcardial perfusion with 4% paraformaldehyde) was co-stained with MCA-6A9 (1:1000, red) and our CPCA-Calbindin (green). In the stratum radiatum of CA1 region, calretinin expresses in a small number of interneurons that do not express calbindin. As a result, our antibodies label different neurons in either red or green. Insets are high-magnification images of the boxed area in each picture. Blue is a hoechst DNA staining.

Western blot analysis of MCA-6A9. Blot of rat brain lysates (lane1), recombinant proteins: pavalbumin (lane 2), calretinin (lane 3), calbindin ( lane 4) was probed with MCA-6A9 at 1:5,000. In rat brain lysates, this antibody  recognizes a clean band at 29 kDa which  represents calretinin . Also it reacts with only calretinin, not other calcium-binding proteins.

Product name Anti Calretinin
Description Mouse Monoclonal to calretinin.
Reference Code MCA-6A9
HGNC name CALB2
RRID# AB_2572243
Molecular weight 29 kDa
Immunogen Full-length recombinant human protein
Isotype IgA
Concentration Antibody is supplied as an aliquot of 1 mg/mL of purified antibody  diluted at 50% glycerol/PBS.
Species Reactivity Human, cow, rat, mouse
Applications Western blot, ICC/IF, IHC
Suggestions for use Western blots: 1:2,000-5,000.
ICC/IF or IHC: 1:2,000-5,000

Calretinin, first described in 1987, acquired its name based on homology with calcium binding protein, calbindin D28k and the tissure of first detection ( Chick retina) (1). As a member of the large superfamily of cytoplasmic Ca2+ binding proteins, Calretinin belongs to the subclass of these containing the “EF hand” Ca2+ binding motif originally characterized in parvalbumin (2). Calretinin is expressed in mammalian central nerve system, testis, fallopian tube and pancreas. In the brain it is localized in certain classes of neurons, and antibodies to it are useful for identifying specific neuronal cell types (3). It is particularly concentrated in granular cells and their parallel fibres, but is also found in many GABAergic interneurons in the cortex. These GABAergic interneurons in most cases express only one of three Ca2+ binding proteins, namely calretinin, calbindin or parvalbumin . As a result these important inhibitory interneurons can be identified and subclassified based on their content of these three proteins (3). Each type of neuron as defined in this fashion has particular electrophysiological and functional properties. For example, calbindin positive interneurons are not fast-spiking as are parvalbumin expressing interneurons. Human calretinin is also known as 29 kDa calbindin and calbindin-2, for its sequence is related to calbindin. Calretinin contains six EF-hand domains. Four of them bind Ca2+ with high affinity in a cooperative manner, one with low affinity and the last one is non-functional, without Ca2+-binding ability (4,5). The function of calretinin appears to be primarily buffering the Ca2+ level in cells and affect intracellular calcium signals. Calretinin deficiency in mossy cells of dentate gyrus and granule cells results abnormal excitability in the cerebellar neuronal network and impairment of long-term potentiation and motor coordination (6). The HGNC name for this protein is CALB2. This antibody was raised against human calretinin protein expressed in and purified from E.coli.


References:

1: Rogers JH: Calretinin: a gene for a novel calcium-binding protein expressed principally in neurons. J Cell Biol 105:1343-1353 (1987).

2: Kretsinger RH & Nockolds CE. Carp Muscle Calcium-binding Protein: II. Structure determination and general description.J. Biol. Chem. 248:3313-3326 (1973).

3: Andressen C, Bliimcke I & Celio MR. Calcium-binding proteins: selective markers of nerve cells. Cell Tissue Res 271:181-208 (1993).

4: Schwaller B, Durussel I, Jermann D, Herrmann B, Cox JA: Comparison of the Ca2+-binding properties of human recombinant calretinin-22k and calretinin.J Biol Chem 272: 29663-29671 (1997).

5: Stevens J, Rogers JH: Chick calretinin: purification, composition, and metal binding activity of native and recombinant forms. Protein Expr Purif 9: 171-181, (1997).

6: Schiffmann SN, Cheron G, Lohof A, d’Alcantara P, Meyer M, Parmentier M, Schurmans S. Impaired motor coordination and Purkinje cell excitability in mice lacking calretinin. Proc Natl Acad Sci U S A. 27: 5257-62 (1999).

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