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CaV1.2 (CACNA1C) Channel Overexpressing Membrane Fraction

A Positive Control for Detecting CaV1.2 (CACNA1C) Channels in Western Blot Analysis
Cat #: LX-104
Lyophilized Powder yes
Accession (Uniprot) Number M57974.1
Origin Rabbit CaV1.2 cDNA (M57974.1) was transcribed in vitro to produce full length mRNA. CaV1.2 mRNA was injected into Xenopus oocytes and protein expression was verified by detecting voltage dependent Nifedipine (#N-120) sensitive ionic current. Membrane fraction was prepared from oocyte population homogenate and lyophilized. Non-injected oocytes underwent the same procedure excluding mRNA injection.
Storage before reconstitution Lyophilized membrane fraction products can be stored at room temperature for up to 2 weeks. For longer periods they should be stored at -20°C.
Reconstitution This product includes:
1 x 0.2 ml lyophilized CaV1.2 (CACNA1C) channel overexpressing Xenopus oocytes membrane fraction
1 x 0.2 ml lyophilized non-injected Xenopus oocytes membrane fraction

Centrifuge the vials containing lyophilized material at 10,000 x g for 5 min before opening and handling them. It’s recommended to work on ice. To reconstitute the product, add 200 µl of Sample Buffer X1 to each vial and carefully pipette to create a homogeneous solution of approximately 1 µg/µl total protein.
It is also possible to reconstitute by first adding 100 µl ddH2O and then 100 µl of Sample Buffer X2.
Heat samples at 70°C for 5min before loading.
Storage after reconstitution Up to 2 weeks at -20°C. Avoid repeated thawing and freezing.
Western blot
Note

Lyophilized membrane fraction was reconstituted in 100 μl ddH2O. Then, 100 μl of Sample Buffer X2 was added to create a loading sample containing approximately 1 μg/μl total protein in final volume of 200 μl. Heated samples (70°C, 5 min) were loaded onto 6% acrylamide gel (45 μl/lane) and resolved using SDS-PAGE, followed by Western Blot analysis using anti-CaV1.2 antibodies Anti-CaV1.2 (CACNA1C) Antibody (#ACC-003), Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-CaV1.2a (CACNA1C) Antibody (#ACC-013), according to product instructions.

Scientific background

All L-type calcium channels are encoded by one of the CaV1 channel genes. These channels play a major role as a Ca2+ entry pathway in skeletal, cardiac and smooth muscles as well as in neurons, endocrine cells and possibly in non-excitable cells such as hematopoetic and epithelial cells. All CaV1 channels are influenced by dihydropyridines (DHP) and are also referred to as DHP receptors.

While the CaV1.1 and CaV1.4 isoforms are expressed in restricted tissues (skeletal muscle and retina, respectively), the expression of CaV1.2 is ubiquitous and CaV1.3 channels are found in the heart, brain and pancreas. Several peptidyl toxins are described that are specific L-type channel blockers, but so far no selective blocker for one of the CaV1 isoforms have been described. These include the Mamba toxins Calcicludine (#SPC-650), Calciseptine (#C-500) and FS-2 (#F-700).

Last update: 23/08/2020

CaV1.2 (CACNA1C) Channel Overexpressing Membrane Fraction
A Positive Control for Detecting CaV1.2 (CACNA1C) Channels in Western Blot Analysis

This product includes:
1 x 0.2 ml lyophilized CaV1.2 (CACNA1C) channel overexpressing Xenopus oocytes membrane fraction
1 x 0.2 ml lyophilized non-injected Xenopus oocytes membrane fraction

CaV1.2 (CACNA1C) Channel Overexpressing Membrane Fraction (#LX-104) is a lyophilized membrane fraction homogenate of Xenopus oocytes that overexpress the CaV1.2 channel. CaV1.2 channels functional and robust expression on membrane surface is confirmed electrophysiologically and pharmacologically prior to membrane fraction isolation.
CaV1.2 (CACNA1C) Channel Overexpressing Membrane Fraction is your positive control for validating Alomone Labs Anti-CaV1.2 (CACNA1C) Antibody (#ACC-003), Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-CaV1.2a (CACNA1C) Antibody (#ACC-013) antibodies. Non-injected Xenopus oocytes membrane fraction supplied in this product serves as negative control.

CaV1.2 (CACNA1C) Channel Overexpressing Membrane Fraction product is:
✓ Lyophilized powder
✓ Economical
✓ Shipped at room temperature (no need for dry ice and extra shipping costs)
✓ User-friendly & time-saving

CaV1.2 (CACNA1C) Channel Overexpressing Membrane Fraction, along with Anti-CaV1.2 (CACNA1C) Antibody, Guinea pig Anti-CaV1.2 (CACNA1C) Antibody and Anti-CaV1.2a (CACNA1C) Antibody, are available as part of the following kits:
CaV1.2 (CACNA1C) Channel Antibody and Membrane Fractions Kit (#LK-104A)
CaV1.2a (CACNA1C) Channel Antibody and Membrane Fractions Kit (#LK-104B)
CaV1.2 (CACNA1C) Channel Antibody and Membrane Fractions Kit (#LK-104C)

For research purposes only, not for human use
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