681507 Sigma-AldrichAnti-Ca_V1.2 (818-835) Rabbit pAb

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision	09-March-2021 JSW
Synonyms	Anti-L-Type Ca2+ Channel
Application	Immunoblotting (1:200) Immunocytochemistry (see application references) Immunoprecipitation (see application references) Immunohistochemistry (!:100)
Application Data	Detection of rat CaV1.2 by immunoblotting. Samples: Extract from rat brain membranes left untreated (lane 1) and treated by preincubation with the peptide antigen (lane 2). Primary antibody: Anti-CaV1.2 (818-835) Rabbit pAb (Cat. No. 681507) (1:200). Detection: chemiluminescence. Detection of mouse CaV1.2 by immunohistochemistry. Samples: Mouse cerebellum with CaV1.2 channel (red) appearing in Purkinje cells (horizontal arrows) and is distributed diffusely in the molecular layer (Mol) including in Purkinje dendrites (vertical arrows) (Figure A). Staining of Purkinje nerve cells with mouse anti-calcium binding protein antibody (green) in the section demonstrates the location of dendrites in the molecular layer (Figure B). Confocal merge of CaV1.2 and CBD28K (Figure C). Primary antibody: Anti-CaV1.2 (818-835) Rabbit pAb (Cat. No. 681507) (1:100). Detection: fluorescence.
Description	Immunoaffinity purified Rabbit polyclonal antibody. Recognizes the ~190 kDa and the ~210 kDa forms of the α1C-subunit of the voltage-gated Ca2+ channel. Supplied with a control peptide (immunogen).
Host	Rabbit
Immunogen species	Rat
Immunogen	a synthetic peptide [(C)TTKINMDDLQPSENEDKS] corresponding to amino acids 848-865 of α1C-subunit of rat brain voltage-gated calcium channels, conjugated to KLH
Isotype	IgG
Species	mouse, rabbit, rat
Positive control	Rat brain or heart
Form	Lyophilized
Formulation	50 µl antibody lyophilized from PBS, pH 7.4, 1% BSA, 0.05% NaN₃ and 40 µg lyophilized control peptide (immunogen).
Preservative	None
Comments	For use as a negative control, preincubate 1 µg of CaV1.2 control peptide with 1 µg of antibody for 1 h at room temperature. Caution: α1-subunits of voltage-gated Ca2+ channels are highly sensitive to proteases. All procedures involving a full-length protein should be performed at 4°C and include the following protease inhibitor cocktail: pepstatin A (Cat. No. 516482; 1 µg/ml), leupeptin (Cat. No. 108975; 1 µg/ml), aprotinin (Cat. No. 616398; 1 µg/ml), benzamidine (Cat. No. 199001; 0.1 µg/ml), AEBSF (Cat. No. 101500; 0.2 mM) and calpain inhibitors I (LLNL, Cat. No. 208719; 8 µg/ml) and II (ALLM, Cat. No. 208721; 8 µg/ml). Variables associated with assay conditions will dictate the optimal working dilution.
Storage	-20°C
Do Not Freeze	Ok to freeze
Special Instructions	Dilutions of the antibody should be made in a physiological buffer containing a carrier protein such as 1-3% BSA. Reconstitute the CaV1.2 control peptide with 100 µl deionized H₂O, aliquot and freeze (-20°C). Avoid freeze/thaw cycles of antibody and peptide solutions. To reduce nonspecific staining, centrifuge all antibody preparations before use (10,000 x g for 5 min) to remove any aggregates.
Toxicity	Standard Handling
References	Hell, J.W., et al. 1995. EMBO J. 14, 3036. Westenbroek, R.E., et al. 1995. J. Neurosci. 15, 6403. Hell, J.W., et al. 1993. J. Cell Biol. 123, 949. Dubel, S.J., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 5058. Snutch, T.P. et al. 1991. Neuron 7, 45.
Application references	Immunocytochemistry Hell, J.W., et al. 1995. EMBO J. 14, 3036. Immunoprecipitation Hell, J.W., et al. 1995. EMBO J. 14, 3036. Dubel, S.J., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 5058.