L-Type Ca Channel antibody

Overview


	Antigen:	L-Type Ca Channel
	Application:	Immunoprecipitation (IP)
	Host:	Mouse
	Quantity:	100 µg (1 mg/mL)
	Price:	241,00 € (Plus shipping costs and VAT)
	Order number:	ABIN130675
	Availability:	7 to 10 days until shipment

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Additional Information: L-Type Ca Channel antibody

Antigen	L-Type Ca Channel
Immunogen	This antibody was purified from hybridoma (clone 7H7) supernatant using protein A agarose. This hybridoma was established by fusion of mouse myeloma cell NS-1 with Balb/c mouse splenocyte immunized with rabbit recombinant cardiac L type Ca 2+ channel a1 subunit C terminal (549a.a.).
Antibody Type
Isotype	IgG2a »Matching secondary antibodies
Description	Voltage-gated calcium channels (VGCCs) mediate Ca2+ entry into cells in response to membrane depolarization. VGCCs are thus involved in a variety of calcium-dependent processes such as myocardial and vascular smooth muscle contraction, neurotransmitter release, hormone secretion, and cell metabolism. Ca2+ entry through the cardiac VGCC determines the rate and force of contraction, and modulation of Ca2+ channel activity contributes to physiological regulation of cardiac function by the sympathetic nervous system. In addition, augmentation of Ca2+ entry through the Long-lasting (L-type) VGCCs is one means by which ß-adrenergic receptor stimulation increases the inotropic state of the myocyte. L-type VGCCs have 5 subunits, a 1, a2, ß, ., and d, and regulation of the channel appears to be controlled primarily by interaction of various cellular proteins or chemical antagonists with the a1 subunit. Modification of calcium channels has been associated with cardiovascular and neurodegenerative diseases including Parkinson’s, epilepsy, and ALS.
Clone	7H7
Host	Mouse
Specificity	This antibody immunoprecipitates L type Ca 2+ channel a1 subunit (175-190KDa).

Application Details: L-Type Ca Channel antibody

Application	Immunoprecipitation (IP)
Application Notes	Immunoprecipitation 1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50 mM Tris-HCl pH 7.2, 250 mM NaCl, 0.1% NP-40, 2 mM EDTA, 10% glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. 3) Add primary antibody as suggested in the Applications into 200 µL of the supernatant. Mix well and incubate with gentle agitation for 30-120 minutes at 4 o C. Add 20 µL of 50% protein A agarose resuspended in the cold Lysis buffer. Mix well and incubate with gentle agitation for 60 minutes at 4 o C. 4) Wash the agarose 3-5 times with the cold Lysis buffer (centrifuge the tube at 2,500 x g for 10 seconds). 5) Resuspend the agarose in 20 µL of Laemmli’s sample buffer, boil for 3-5 minutes, and centrifuge for 5 minutes. Load 10 µL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 6) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi-dry transfer system (Transfer Buffer - 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for specific transfer procedure. 7) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4 o C. 8) Incubate the membrane with 1:120 anti- L type Ca 2+ channel a1c polyclonal antibody (Alomone, code no. ACC-003) diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. (The optimal antibody concentration will depend on the experimental conditions.) 9) Wash the membrane with PBS (5 minutes x 6 times). 10) Incubate the membrane with the 1:16,000 POD-conjugated anti-rabbit IgG (MBL, code no. 458) diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 11) Wash the membrane with PBS (5 minutes x 6 times). 12) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 13) Expose to X-ray film in a dark room for 5 minutes. Develop the film as usual. The conditions for exposure and development may vary. Positive control for immunoprecipitation, PC12 040716-1 Western blotting: Not recommended. Immunohistochemistry: Not tested. Immunocytochemistry: Not tested. Immunoprecipitation: 2µg / 200µL of cell extract from 5x10 6 cells. Flow cytometry: Not tested.
Concentration	1 mg/mL
Buffer	100 µg IgG in 100 µL volume of PBS containing 50% glycerol, pH 7.2. Contains no preservatives.
Restrictions	For Research Use Only

Images: L-Type Ca Channel antibody

Immunoprecipitation of L type C a2+ channel ?1 subunit from PC 12 cells with mouse IgG 2a(1) or D 155-3 (2). After immunoprecipitated with the antibody,immunocomplex was resolved on SDS-PAGE and immunoblotted with anti-L type Ca2+ channel ?1c polyclonal antibody

Image for L-Type Ca Channel antibody (1)

External Information: L-Type Ca Channel antibody

Google Scholar	Find more references for clone 7H7 on Google Scholar™
EMBL Harvester	Search human, mouse or rat genome for antigen L-Type Ca Channel (Bioinformatic Harvester (EMBL))