Guinea pig Anti-TRPC6 Antibody

Short transient receptor potential channel 6, Transient receptor protein 6, TRP6
    Cat #: AGP-002
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Guinea pig
    Reactivity: m, r
    May also work in: h
    Immunogen
    Peptide (C)RRNESQDYLLMDELG, corresponding to amino acid residues 24-38 of mouse TRPC6 (Accession Q61143). Intracellular, N-terminus.
    Accession (Uniprot) Number Q61143
    Gene ID 22068
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat - identical; human - 15/16 amino acid residues identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
    Antibody concentration after reconstitution 0.9 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
    Control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ih, wb
    May also work in: ic, ifc, ip
    Western blot
    Western blot analysis of rat brain membranes:
    1. Guinea pig Anti-TRPC6 Antibody (#AGP-002), (1:200).
    2. Guinea pig Anti-TRPC6 Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of TRPC6 in rat cerebellum
    Immunohistochemical staining of rat cerebellum using Guinea pig Anti-TRPC6 Antibody (#AGP-002). A. TRPC6 (green) appears in the soma of Purkinje cells (white arrow). B. Parvalbumin (red), a marker of Purkinje and interneuronal cells, is stained in the same section. C. Merge of the images demonstrates intense expression of TRPC6 in the soma of Purkinje cells and lower expression in the molecular (Mol) layer. DAPI is used as the counterstain (blue).
    Immuno-colocalization of TRPC6 and TRPC3 in rat cerebellum
    Immunohistochemical staining of rat cerebellum frozen section using Guinea pig Anti-TRPC6 Antibody (#AGP-002) and rabbit Anti-TRPC3-ATTO-594 Antibody (#ACC-016-AR). A. TRPC6 staining (green) appears in molecular layer and in Purkinje cells. B. In the same section as in A, staining of TRPC3 (red) appears as well in both molecular layer and Purkinje cells. C. Merge images of A and B indicates co-localization in Purkinje cells and molecular layer.
    Immuno-colocalization of TRPC6 and TRPC3 in rat cerebellum
    Immunohistochemical staining of rat cerebellum frozen section using Guinea pig Anti-TRPC6 Antibody (#AGP-002) and rabbit Anti-TRPC3 Antibody (#ACC-016). A. TRPC6 staining (green) appears in molecular layer and in Purkinje cells. B. In the same section, staining of TRPC3 (red) appears as well in both molecular layer and Purkinje cells. C. Merge images of A and B indicates extensive co-localization. DAPI is used as the counterstain (blue).
    References
    1. Ramsey, S.I. et al. (2006) Annu. Rev. Physiol. 68, 619.
    2. Pedersen, S.F. et al. (2005) Cell. Calcium. 38, 233.
    3. Montell, C. (2005) Sci. STKE 2005, re3.
    4. Hilgemann, D.W. et al. (2001) Sci. STKE 2001, re19.
    5. Abramowitz, J. et al. (2009) FASEB J. 23, 297.
    6. Winn, M.P. et al. (2005) Science308, 1801.
    7. Reiser, J. et al. (2005) Nat. Genet37, 739.
    Scientific background

    Transient receptor potential (TRP) channels are relatively non-selective ion channels enabling the exchange of cations down their electrochemical gradient. This exchange enables the intracellular rise in Na+ and Ca2+ concentration and ultimately in the cell membrane depolarization, important for action potential propagation and muscle contraction1. They are activated by an extremely broad range of stimuli namely, temperature, voltage, pH, endocrine factors as well as signaling molecules2.

    The TRP channel family is composed of 28 members divided in 7 subgroups: TRPV, TRPC, TRPM, TRPA, TRPN, TRPP and TRPML. All members of the TRP family form tetramers and could heterhomultimerize. They have 6 transmembrane (TM) domains, and a pore domain between the fifth (S5) and sixth (S6) transmembrane domains. In general, TRP channels enable the passage of either Na+ or Ca2+ ions with little or no preference. However, some channels do exhibit some selectivity. Also, TRP channels do not display the positive charges in the S4 voltage-sensing domain like most voltage sensitive channels, although they do display voltage dependency3. In addition, TRP channels have in the C-terminal intracellular region to the S6 domain a TRP domain comprising 25 amino acids that is more or less conserved among most TRP channels. Within the TRP domain, there is a TRP box composed of six amino acids, and TRP box 2 – a proline rich domain1,3. The TRP domain seems to be responsible for the binding of PIP2, a phospholipid important for the regulation of channel activity4.

    The TRPC subfamily is further divided into the following: TRPC1/4/5, TRPC3/6/7 and TRPC21. Activation of phospholipase C (PLC) ultimately leads to the formation of diacylglycerol (DAG) and inositol 1,4,5-triphosphate (IP3) via hydrolysis of PIP2. The increase in concentration of these intracellular second messengers leads to the activation of non-selective Ca2+ channels and an IP3-induced release of Ca2+ from intracellular stores5. The intracellular Ca2+ store depletion in turn activates Ca2+ specific channels to allow replenish intracellular Ca2+ levels. TRPCs are thought to be activated upon intracellular store Ca2+ depletion, and may function in concert with the recently identified Orai channel5. The TRPC3/6/7 class produces similar currents upon activation.

    TRPC6 can form heterotetramers with TRPC3 and TRPC7. It is primarily expressed in brain, lung and muscle. High levels of expression of the channel were also found in human platelets. Recently it was reported that TRPC6 is also expressed in the kidney where a mutated channel has been implicated in kidney failure disease6,7.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Last update: 22/11/2018

    Guinea pig Anti-TRPC6 Antibody (#AGP-002), raised in guinea pigs, is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize TRPC6 from mouse, rat, and human samples. The antigen used to immunize guinea pigs is the same as Anti-TRPC6 Antibody (#ACC-017) raised in rabbit. Our line of guinea pig antibodies enables more flexibility with our products such as immuno-colocalization studies, immunoprecipitation, etc.

    For research purposes only, not for human use