Monoclonal anti parvalbumin

For long storage, keep small aliquots of 1-2 µl at - 80°C (or at least - 20°C). For continous use keep diluted solutions at 4°C (with 0.01% Na-azide). Avoid repeated freezing and thawing.

Monoclonal anti-Parvalbumin is a mouse IgG1 produced by hybridization of mouse myeloma cells with spleen cells from mice immunized with parvalbumin purified from carp muscles (1). The antibody was evaluated for specificity and potency: a) by indirect immunofluorescent or immunoperoxidase labeling as well as Avidin-Biotin staining of cryostat or vibratome-sections of 4% paraformaldehyde fixed tissue; b) by immunoenzymatic labelling of immunoblots; c) by radioimmunoassay (RIA)(1). The product is a monoclonal antibody (McAB) against parvalbumin, a calcium-binding protein of the EF-hand family related to calmodulin and troponin-C. The antibody reacts specifically with parvalbumin in tissue originating from human, monkey, rabbit, rat, mouse chicken and fish. The McAB specifically stains the 45Ca-binding spot of parvalbumin (MW 12'000 and IEF 4.9) in a two-dimensional "immunoblot". In a RIA set up the McAB measures parvalbumin with a sensitivity of 10 ng/assay and an affinity of 7.9 x 1012 L/M (1).

Parvalbumin

Mouse monoclonal, IgG1

Lyophilized ascites (no preservatives).

1. Celio M.R. et al., Cell Calcium 9:81-86, 1988 2. Celio M.R., Heizmann C.W. (1981) Nature 293: 300-302 3. Celio M.R., Heizmann C.W. (1982) Nature 297:504-506 4. Celio M.R. (1986) Science 231:995-997 5. Schwaller B., et al. (1999) Am. J. Physiol. 276. C395-403

Immunohistochemistry: 1:5'000 - 1:10'000 Immunoblotting: 1:1'000. We recommend that the optimal dilution be determined by titration test.

Lot No: 10-11 (F) Reconstitution: with 200 µl bidistilled water. Immunoblot: The specificity of antibody 235 could be determined by immunoblots of brain and muscle extracts (1). However, this antibody does not recognize well the parvalbumin molecule after SDS-gel electrophoresis. Immunohistochemistry on Calbindin D-28k knock-out mice: Antibody 235 immunolabels a subpopulation of neurons in the normal brain with high efficiency (Fig. 2a), but does not stain the brain of parvalbumin knock out mice (Fig. 2b). Product: Conc. supernat. uses: Calcium binding-proteins represent a family of small, acidic proteins equipped with peculiar cavities which accept Ca2+ with high selectivity. There are two types of calcium binding-proteins, “trigger” and “buffer” proteins. Those of the “trigger”-type (e.g. calmodulin and troponin-C) act by changing shape upon binding Ca2+. This distortion exposes regions on the surface of the protein, which interact with surrounding target molecules, altering their activity. The calcium binding-protein of the “buffer”-type are conceived as a system which is in charge of controlling the Ca2+ concentration inside cells. Parvalbumin occurs mainly in subpopulations of nerve cells (2,4) and in fast muscle fibers (3). It might confer on these cells peculiar skills in the handling of calcium-ions. Parvalbumin is an attractive neuroanatomical tool and this McAB guarantees the continued production of a constant titer of anti-parvalbumin antibody with the same specificity and chemical identity. Antibody performance: McAB 235 against parvalbumin specifically localizes parvalbumin using free-floating or mounted sections of brain and muscles probably of all vertebrates. Fixation does not seem to be an hindrance to the detection of the antigen since even 2.5% glutaraldehyde was compatible with immunolocalization on semithin (0.5 µm thin) cryo-sections of the rat cortex (4). It can be used with rat-parvalbumin as a tracer to quantify parvalbumin in the brain of various species (1).