KV1.2 (KCNA2) Channel Overexpressed Membrane Fractions

A Positive Control for Detecting KV1.2 (KCNA2) Channels in Western Blot Analysis
    Cat #: LX-101
  • Lyophilized Powder
  • Accession (Uniprot) Number NM_012970
    Gene ID 25468
    Origin Rat KV1.2 cDNA (NM_012970) was transcribed in vitro to produce full length mRNA. KV1.2 mRNA was injected into Xenopus oocytes and protein expression was verified by detection of voltage evoked Tityustoxin Kα (#STT-360) 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 injected and non-injected membrane lysates can be stored at room temperature for 2 weeks. For longer periods, they should be stored at -20°C.
    Reconstitution Work on ice. Gently reconstitute injected and non-injected membrane lysates in 100 μl DDW (total protein concentration is 0.4 μg/μl). Centrifuge all product preparations before use (10000 x g 5 min).
    Storage after reconstitution Up to 2 weeks at -20°C. Avoid repeated thawing and freezing.
      • KV1.2 (KCNA2) Channel Overexpressed Membrane Fractions
        Western blot analysis of KV1.2 (KCNA2) Channel Overexpressed Membrane Fractions (#LX-101) obtained from Xenopus oocyte membrane fractions expressing rat KV1.2 channels (lanes 2 and 4), and non-injected oocyte lysates (lanes 1 and 3):
        1,2. Anti-KV1.2 (KCNA2) Antibody (#APC-010), (1:200).
        3,4. Anti-KV1.2 (KCNA2) Antibody, preincubated with the negative control antigen.
    Note

    Lyophilized membrane fraction was reconstituted in 100 μl DDW (see below). 100 μl of Sample Buffer 2X was added (total protein concentration was 0.2 μg/μl). Boiled samples were loaded onto 10% acrylamide gel (40-50 μl/lane) and resolved using SDS-PAGE gel, followed by western blot analysis using Anti-KV1.2 (KCNA2) Antibody (#APC-010) following product instructions.

    This product includes:
    1 x 0.1 ml lyophilized KV1.2 channel overexpressed membrane lysate
    1 x 0.1 ml lyophilized non-injected Xenopus oocyte membrane lysate

    References
      • KV1.1 is a mammalian voltage-dependent K+ channel, homologous to the Drosophila Shaker K+ channel. KV1.1 was the first mammalian KV channel to be cloned from mouse brain.1 Eight Shaker-related genes exist in mammals constituting the KV1, subfamily of the large KV channel family of genes.2

        A functional KV1 channel is either a membrane spanning homotetramer or heterotetramer, which is composed of members of the same subfamily. In addition several auxiliary subunits and intracellular proteins might interact with the channel and affect its function.

        The structure of KV1.1 channel is similar to all KV channels and includes six membrane spanning helixes creating a voltage sensor domain and a pore domain.2

        The channel is expressed in neurons and cardiac and skeletal muscle tissue as well as in retina and pancreas.2 The functional channel is considered low voltage activated and shows very little inactivation. Therefore, this channel activity influences the membrane potential and excitability of neurons and muscle. Mutations in the coding of KV1.1 gene were discovered in Episodic Ataxia patients.3

        KV1.1 channels are sensitive to low doses of TEA (0.3 mM) and 4-AP (0.29 mM), the “classical” non-selective potassium channel blockers.

        Several venomous toxins from snakes, scorpions and sea anemones are potent blockers (affecting the channels in the nanomolar range) of KV1.1 channels. Among these the most potent and selective are α-Dendrotoxin (#D-350, 0.4-4 nM) and δ-Dendrotoxin (#D-380, 0.03-1.8 nM), Dendrotoxin-K (#D-400, 0.03 nM), Agitoxin-2 (#STA-420, 0.044 nM) and Hongotoxin-1 (#STH-400, 0.031 nM).4

    Last update: 13/01/2020

    KV1.2 (KCNA2) Channel Overexpressed Membrane Fractions (#LX-101) are Xenopus oocyte membrane fractions overexpressing the KV1.2 channel.* KV1.2 (KCNA2) Channel Overexpressed Membrane Fractions are your positive control for validating Alomone Labs Anti-KV1.2 (KCNA2) Antibody (#APC-010).

    Overexpressed Membrane Fractions are:
    ✓ Lyophilized powder
    ✓ Economical
    ✓ Shipped at room temperature (no need for dry ice and extra shipping costs)
    ✓ User-friendly & time-saving. Just add water, sample buffer and load your gel

    KV1.2 (KCNA2) Channel Overexpressed Membrane Fractions, along with Anti-KV1.2 (KCNA2) Antibody, are available as part of the following kit:
    KV1.2 (KCNA2) Channel Antibody and Membrane Fractions Kit (#LK-101).

    *Product includes non-injected oocyte membrane lysates

    For research purposes only, not for human use