Anti-Capsaicin receptor (VR1/TrpV1) Antibody (BS397)

Name: Anti-Capsaicin receptor (VR1/TrpV1) Antibody
Brand: Biosensis
SKU: M-1714-100
Price: 337.0 USD
Availability: InStock

Our Anti-Capsaicin receptor (TrpV1) mouse monoclonal primary antibody detects guinea pig (predicted), mouse, and rat Capsaicin receptor (TrpV1), and is IgG. It is validated for use in FC, WB.

A: Analysis of TRPV1 expression in rat PC12 cell line by Flow Cytometry. Fixing and permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 1% BSA, Primary antibody: Mouse Monoclonal antibody to TRPV1 (cat # M-1714-100, 2 µg per ~10^6 cells) for 30 minutes at RT, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 dilution), 20 minutes in dark at room temperature. Negative control: Non-specific Control IgG, clone X63 (cat # M-1249-200, black dashed). Data and results were generated using Orflo MoxiflowTM instrument and protocols.B: Western blot of TrpV1 in rat PC12 cell lysates (80 µg/lane). M-1714-100 detects TrpV1 protein at 95-100 kDa. SDS-PAGE: denatured and reduced; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 µm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C (2 µg/mL); Secondary antibody: anti-mouse-HRP (1/6000) 2 hours at RT; Detection: Chemiluminiscence.C: Immunohistochemical staining of TrpV1 in mouse dorsal root ganglia. Immunoreactivity was visualized with anti-mouse-Cy3 conjugate (red). Magnification: 20x. Courtesy P. Vilimas, Flinders University Adelaide.D: Western blot (denatured and reduced) of TrpV1 in cell lysates of forskolin and NGF stimulated 50B11 hybrid mouse x rat DRG cell lines and NGF-stimulated PC12 cells (10 µg/lane). M-1714-100 detects monomeric TrpV1 protein at 95-100 kDa. Primary antibody: 1 µg/mL (4°C overnight). Detection: Chemiluminiscence. Courtesy Dr. D. Matusica, Flinders University.

A: Immunofluorescence analysis of TrpV1 expression in rat C6 glioma cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) C6 cells were incubated with TrpV1 antibody M-1714-100 (clone BS397, 2 µg/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (5 µg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). Magnification: 100x.B: Analysis of TRPV1 expression in C6 cells by Flow Cytometry. Fixing and permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 200 µg/mL sheep IgG, Primary antibody: Mouse Monoclonal antibody to TRPV1 (cat # M-1714-100, 2 µg per ~106 cells) for 30 minutes at room temperature, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 dilution), 20 minutes in dark at room temperature. Negative control: Non-specific Control IgG, clone X63 (cat # M-1249-100, black dashed). Data and results were generated using Orflo MoxiflowTM instrument and protocols.

Western blot of TrpV1 in mouse brain homogenate (25 µg/lane). M-1714-100 detects bands at 90-100 kDa and 180-200 kDa corresponding to capsaicin receptor monomer and dimer. SDS-PAGE: denatured and reduced; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 µm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C; Secondary antibody: anti-mouse-HRP (1/6000) 2 hours at RT; Detection: Chemiluminiscence.

A: Analysis of TRPV1 expression in rat PC12 cell line by Flow Cytometry. Fixing and permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 1% BSA, Primary antibody: Mouse Monoclonal antibody to TRPV1 (cat # M-1714-100, 2 µg per ~10^6 cells) for 30 minutes at RT, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 dilution), 20 minutes in dark at room temperature. Negative control: Non-specific Control IgG, clone X63 (cat # M-1249-200, black dashed). Data and results were generated using Orflo MoxiflowTM instrument and protocols.
B: Western blot of TrpV1 in rat PC12 cell lysates (80 µg/lane). M-1714-100 detects TrpV1 protein at 95-100 kDa. SDS-PAGE: denatured and reduced; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 µm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C (2 µg/mL); Secondary antibody: anti-mouse-HRP (1/6000) 2 hours at RT; Detection: Chemiluminiscence.
C: Immunohistochemical staining of TrpV1 in mouse dorsal root ganglia. Immunoreactivity was visualized with anti-mouse-Cy3 conjugate (red). Magnification: 20x. Courtesy P. Vilimas, Flinders University Adelaide.
D: Western blot (denatured and reduced) of TrpV1 in cell lysates of forskolin and NGF stimulated 50B11 hybrid mouse x rat DRG cell lines and NGF-stimulated PC12 cells (10 µg/lane). M-1714-100 detects monomeric TrpV1 protein at 95-100 kDa. Primary antibody: 1 µg/mL (4°C overnight). Detection: Chemiluminiscence. Courtesy Dr. D. Matusica, Flinders University.

Click on image to zoom

Species Reactivity

Guinea Pig, Mouse, Pig, Rat

Applications

Flow, ICC, IHC, WB

Host Species

Mouse

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Data Sheet

SKU: M-1714-100

Volume/Size: 100 µg

Variant

Price:

$337.00

Quantity:

Ships: 1-2 business days

Product Details

Target

Capsaicin receptor (TrpV1)

Scientific Background

The capsaicin receptor (VR1, TRPV1) is a ligand-activated non-selective calcium permeant cation channel involved in detection of noxious chemical and thermal stimuli. The receptor seems to mediate proton influx and may be involved in intracellular acidosis in nociceptive neurons. It is involved in mediation of inflammatory pain and hyperalgesia. Sensitized by a phosphatidylinositol second messenger system activated by receptor tyrosine kinases, which involves PKC isozymes and PCL. Activation by vanilloids, like capsaicin, and temperatures higher than 42 degrees Celsius, exhibits a time- and Ca2+-dependent outward rectification, followed by a long-lasting refractory state. Mild extracellular acidic pH (6.5) potentiates channel activation by noxious heat and vanilloids, whereas acidic conditions (pH less than 6) directly activate the channel. Can be activated by endogenous compounds, including 12-hydroperoxytetraenoic acid and bradykinin. Acts as ionotropic endocannabinoid receptor with central neuromodulatory effects. Triggers a form of long-term depression (TRPV1-LTD) mediated by the endocannabinoid anandamine in the hippocampus and nucleus accumbens by affecting AMPA receptors endocytosis (Ref: uniprot.org).

Format

IgG

Clonality

Monoclonal

Clone

BS397

Applications

Flow, ICC, IHC, WB

Host Species

Mouse

Immunogen

A synthetic peptide (C-GSLKPEDAEVFKDSMVPGEK) as a part of the C-terminal rat VR1 protein (aa: 819-838) has been used as the immunogen.

Immunogen Species

Rat

Species Reactivity

Guinea Pig, Mouse, Pig, Rat

Storage

Spin vial briefly before opening. Reconstitute in 100 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material. Final buffer contains no preservatives but will contain 0.1% trehalose and buffer salts. Store lyophilized antibody at 2-8°C. After reconstitution divide in to aliquots and store at -20°C for a higher stability. Antibody contains no preservatives. Storage at 2-8°C with an appropriate antibacterial agent. USE Sterile methods. Highest purity Glycerol (1:1) may be added for an additional stability when stored at refrigerated or freezing temperatures. Avoid repetitive freeze/thaw cycles.

Physical State

Lyophilized

Production Notes

Protein G purified mouse immunoglobulin

Buffer

Lyophilized from PBS, pH 7.4 with 0.1% trehalose.

Dilution Ranges

WB: 0.5-2 µg/mL
IHC: 1-10 µg/mL
ICC: 1-2 µg/mL

Application Details

Flow Cytometry: 2 ug/10^6 cells.

Western blotting: 0.5-2 µg/mL , SDS-PAGE on Bis-Tris gel 4-12%, 5% beta-mercaptoethanol, primary antibody O/N incubation in 5% skim milk/TBST. Secondary is anti-mouse-HRP, 1/6000 dilution, 2h at room temperature. Blot developed on Li-Cor C-DiGit Scanner.

IHC: Frozen or PEG embedded tissues tested (PEG embedding, see Klosen P et al (1993) J Histochem Cytochem. 41(3):455-63). Conditions tested: 1-10 µg/mL in PBS, 48 hours, followed by detection via directly conjugated fluorescent anti-mouse secondary.

Immunohistochemistry Protocol for Product Image
4% PFA perfused, formalin fume-fixed DRG tissues: Adult male rat was perfusion-fixed transcardially with 4% paraformaldehyde. The lumbar dorsal root ganglia were isolated and embedded into egg yolk medium, which was solidified in formalin fume. 30-µm-thick floated sections were prepared with a Vibratome. For pretreatment, the sections were thoroughly rinsed in 0.1M PBS (pH 7.6) and then incubated with a mixture of 0.5% H2O2 and 0.2% Triton X-100 for 30 min. Then, antigen retrieval was performed with 0.1M citrate buffer (pH=6.0) at 80°C for 30 min. (Please note that without this step, we did not obtain any labeling). Then, the TRPV1 signal was detected using a 1:5000 dilution of the primary antibody in 2% normal horse serum (NHS) in PBS (16 h; RT), followed by a 1:500 dilution of the biotinylated secondary antibody (2 h in 2% NHS/PBS; RT) and then, the ABC Elite reagent (Vector; 1:1000 in Tris buffer; pH 7.6). The signal was visualized with Ni-DAB chromogen. The immunostained sections were rinsed in Tris buffer, mounted on microscope slides, air dried, counter-stained with cresyl violet, dehydrated and coverslipped with DPX mounting medium.
[Immunohistochemistry protocol generously provided by Dr. Szabolcs Takacs & Dr. Erik Hrabovszky, Institute of Experimental Medicine, Budapest, Hungary].

Antibody not yet tested on paraffin-embedded sections. Other immunohistochemistry methods have not yet been tested but are expected to be reactive under correct conditions. Antigen recovery is suggested; the epitope is internal cytoplasmic and could be covered by internal proteins during fixation.

ICC: 4% formaldehyde fixed cells tested; requires permeabilization step as antigen epitope is intracellular. Suggested primary antibody concentration: 1-2 µg/mL.

Conjugate

Unconjugated

Specificity

Antibody is specific for rat/mouse VR1 protein in Westerns and immunofluorescent immunohistochemistry on mouse PEG fixed DRG tissues. Pre-absorption with immunogen obliterates positive staining. Cross reactivity with other non-VR1 proteins is minimal; cross reactivity with VR1 from other species not yet tested. This antibody clone is known to react with rat and mouse TrpV1. It is predicted to react with guinea pig due to sequence homology.

Usage Statement

For research use only.

Country of Origin

United States

Expiration

12 months after date of receipt (unopened vial).

Synonyms

TRPV1_SON; VR1; Vr1l1; Transient receptor potential cation channel subfamily V member 1; deltaN-TRPV1; TrpV1; vanilloid receptor 1; osm-9-like TRP channel 1; vanilloid receptor type 1 like protein 1; OTRPC1; vanilloid type 1 receptor; Capsaicin receptor; VR-1

UniProt ID

O35433
UniProt Summary: Non-selective calcium permeant cation channel involved in detection of noxious chemical and thermal stimuli. Seems to mediate proton influx and may be involved in intracellular acidosis in nociceptive neurons. Involved in mediation of inflammatory pain and hyperalgesia. Sensitized by a phosphatidylinositol second messenger system activated by receptor tyrosine kinases, which involves PKC isozymes and PCL. Activated by vanilloids, like capsaicin, and temperatures higher than 42 degrees Celsius. Upon activation, exhibits a time- and Ca(2+)-dependent outward rectification, followed by a long-lasting refractory state. Mild extracellular acidic pH (6.5) potentiates channel activation by noxious heat and vanilloids, whereas acidic conditions (pH <6) directly activate the channel. Can be activated by endogenous compounds, including 12-hydroperoxytetraenoic acid and bradykinin. Acts as ionotropic endocannabinoid receptor with central neuromodulatory effects. Triggers a form of long-term depression (TRPV1-LTD) mediated by the endocannabinoid anandamine in the hippocampus and nucleus accumbens by affecting AMPA receptors endocytosis.

NCBI Gene ID

83810

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Anti-Capsaicin receptor (VR1/TrpV1) Antibody (BS397)

SKU: M-1714-100

Product Details

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