Label ATTO-633. Maximum absorption 629 nm; maximum fluorescence 657 nm. The fluorescence is excited most efficiently in the 610 - 645 nm range. This label is analogous to the dyes Alexa 647, Alexa 633 and Cy5 and can be used for direct flow cytometry (FACS) using the He:Ne laser.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 50 µl double distilled water (DDW).
Antibody concentration after reconstitution 1 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, 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 × g 5 min).
Standard quality control of each lot Western blot analysis (unlabeled antibody, #ACC-033), and immunohistochemistry (labeled antibody).
Applications: FC, ICC, IF, IHC, LCI
Immunohistochemistry
Rat dorsal root ganglia (DRG).
Expression of TRPV3 in rat DRG
Immunohistochemical staining of rat dorsal root ganglion (DRG) using Anti-TRPV3 (extracellular)-ATTO Fluor-633Antibody (#ACC-033-FR). A. TRPV3 staining (purple) is detected in DRG neurons. B. Nuclear staining using DAPI as the counterstain (Blue). C. Merge images of A and B.
Direct flow cytometry
Cell surface detection of TRPV3 by direct flow cytometry in live intact human Jurkat T-cell leukemia cell line:
TRP channels are a large family (about 28 genes) of plasma membrane, non-selective cationic channels that are either specifically or ubiquitously expressed in excitable and non-excitable cells.1 The TRP channels have six putative transmembrane domains (TM) with a pore domain between the fifth and the sixth TM, and all assemble as tetramers. Both the N- and the C-terminus of all TRPs are intracellular.3
According to IUPHAR, the TRP family is composed of three main subfamilies on the basis of sequence homology; TRPC, TRPM and TRPV (to date, three additional subfamilies are also considered to belong to the TRP family: the TRPA, TRPML, and TRPP).1-4 The TRPV subfamily consists of six members, TRPV1-6.5
Four members of the TRPV family have been described as a thermosensitive ion channels (TRPV1 to TRPV4). Each channel exhibits distinct thermal activation thresholds ranging from noxious cold (<17°C) to noxious heat (>52°C).6,7
TRPV3 is highly expressed in skin keratinocytes. It forms a nonselective cation channel that is activated by hot temperatures in the innocuous and noxious range. The channel is also expressed in oral and nasal cavities where it has been postulated to play a role in flavor sensation. TRPV3 is also implicated as a molecular target of some allergens and skin sensitizers.8
Application key:
CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot