Overview

  • Product name
  • Description
    Rabbit polyclonal to VR1
  • Host species
    Rabbit
  • Specificity
    This antibody has been optimised on rat tissue. We recommend ab31895 for detection of VR1 in mouse tissue.
  • Tested applications
    Suitable for: WB, IHC-FoFr, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Rat
    Predicted to work with: Mouse, Non human primates
  • Immunogen

    Synthetic peptide:

    RASLDSEESESPPQENSC

    , corresponding to amino-terminus of Rat VR1, amino acids 4-21.

  • Positive control
    • ICC: VR1 transfected cells. WB: dorsal root ganglion cell lysate IHC: dorsal root ganglion (perfusion fixed)
  • General notes


    Capsaicin, the main pungent ingredient in hot chili peppers, elicits a sensation of burning pain by selectively activating sensory neurons that convey information about noxious stimuli to the central nervous system. The protein encoded by this gene is a receptor for capsaicin and is a non-selective cation channel that is structurally related to members of the TRP family of ion channels. This receptor is also activated by increases in temperature in the noxious range, suggesting that it functions as a transducer of painful thermal stimuli in vivo. Four transcript variants encoding the same protein, but with different 5' UTR sequence, have been described for this gene.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.05% Sodium azide
    Constituent: Whole serum
  • Purity
    Whole antiserum
  • Primary antibody notes
    Capsaicin, the main pungent ingredient in hot chili peppers, elicits a sensation of burning pain by selectively activating sensory neurons that convey information about noxious stimuli to the central nervous system. The protein encoded by this gene is a receptor for capsaicin and is a non-selective cation channel that is structurally related to members of the TRP family of ion channels. This receptor is also activated by increases in temperature in the noxious range, suggesting that it functions as a transducer of painful thermal stimuli in vivo. Four transcript variants encoding the same protein, but with different 5' UTR sequence, have been described for this gene.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab10296 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Predicted molecular weight: 100.7 kDa. (see Guo et al reference).
IHC-FoFr 1/1000.

(see Guo et al reference).

ICC/IF 1/1000.
IP Use at an assay dependent concentration. PubMed: 25151644

Target

  • Function
    Receptor-activated 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. May be 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. 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 accubens by affecting AMPA receptors endocytosis.
  • Tissue specificity
    Widely expressed at low levels. Expression is elevated in dorsal root ganglia. In skin, expressed in cutaneous sensory nerve fibers, mast cells, epidermal keratinocytes, dermal blood vessels, the inner root sheet and the infundibulum of hair follicles, differentiated sebocytes, sweat gland ducts, and the secretory portion of eccrine sweat glands (at protein level).
  • Sequence similarities
    Belongs to the transient receptor (TC 1.A.4) family. TrpV subfamily. TRPV1 sub-subfamily.
    Contains 6 ANK repeats.
  • Domain
    The association domain (AD) is necessary for self-association.
  • Post-translational
    modifications
    Phosphorylation by PKA reverses capsaicin-induced dephosphorylation at multiple sites, probably including Ser-117 as a major phosphorylation site. Phoshphorylation by CAMKII seems to regulate binding to vanilloids. Phosphorylated and modulated by PKCM and probably PKCZ. Dephosphorylation by calcineurin seems to lead to receptor desensitization and phosphorylation by CAMKII recovers activity.
  • Cellular localization
    Cell junction > synapse > postsynaptic cell membrane. Cell projection > dendritic spine membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Capsaicin receptor antibody
    • DKFZp434K0220 antibody
    • osm 9 like TRP channel 1 antibody
    • Osm-9-like TRP channel 1 antibody
    • OTRPC1 antibody
    • Transient receptor potential cation channel subfamily V member 1 antibody
    • TRPV 1 antibody
    • Trpv1 antibody
    • TRPV1_HUMAN antibody
    • Vanilloid receptor 1 antibody
    • Vanilloid receptor subtype 1 antibody
    • VR 1 antibody
    • VR1 antibody
    see all

Images

  • ICC/IF image of ab10296 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab10296, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • VR1 staining in the rat dorsal root ganglion using ab10296.

References

This product has been referenced in:
  • Hulse RP  et al. Regulation of alternative VEGF-A mRNA splicing is a therapeutic target for analgesia. Neurobiol Dis 71:245-59 (2014). IP ; Mouse . Read more (PubMed: 25151644) »
  • van den Wijngaard RM  et al. Essential role for TRPV1 in stress-induced (mast cell-dependent) colonic hypersensitivity in maternally separated rats. Neurogastroenterol Motil 21:1107-e94 (2009). IHC-FoFr ; Rat . Read more (PubMed: 19523146) »
See all 4 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

I'm very sorry to hear you are experiencing problems with ab10296. While this antibody has not been tested in horse, it should work very well in western blotting in rat DRG lysate. I think the problem may lie in two steps: 1) the extraction of VR1 from the lysates. We are not familiar with the Tri Reagent from Molecular Research Center and we recommend the following lysis buffer: 10mM Tris buffer pH 7.5 containing 0.16M sucrose, 1mM EDTA, 1mM DTT and protease inhibitors. Please omit the solubilisation step and simply leave the homogenate to rotate at 4C for 2 hours, then centrifuge the samples and collect the supernatant, then immediately boil in reducing loading buffer. 2) the antibody does not have time to bind efficiently to the protein on the blot. Please try incubating the antibody overnight at 4C in TBST , you may need to try 1:500 but I think already incubating longer will give you a better result. I think the following small changes will also make a difference: -load more lysate per well (typical amounts are 20-40ug) -block in 5%BSA for 1 hour and incubate the antibody in TBST only or 1% BSA in TBST. Sometimes milk can prevent the antibody from binding to the membrane, unfortunately. -please check that the secondary works well with other primary antibodies. I hope these recommendations will resolve your problem. Please do not hesitate to contact me if you still experience problems with those changes, I would be happy to offer you a replacement vial or refund if the antibody was purchased in the last 120 days.

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Answer

Thanks for your enquiry and I'm sorry to hear that your customer is experiencing difficulty with their latest vial of ab10296. We haven't received any other complaints regarding this batch, but I would be happy to provide a free of charge replacement vial. Lot 160975 is no longer in stock and I will have a vial from a newer lot sent to you on your next shipment. Please let me know if you have any additional questions or concerns.

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Answer

Thank you for your enquiry. Following correspondence with the source of this antibody I have determined that it has been used and characterized by western blotting in the following publication: Guo A, Vulchanova L, Wang J, Li X, Elde R. (1999) Immunocytochemical localization of the vanilloid receptor 1 (VR1): relationship to neuropeptides, the P2X3 purinoceptor and IB4 binding sites. Eur J Neurosci. 11(3):946-58. PMID: 10103088 Following examination of this publication I can tell you that this antibody produces "a broad protein band with molecular weight around 95-100KDa". I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

Thank you for you enquiry. We would not recommend fixing in formalin for ICC. We would recommend ethanol, methanol or ICE cold acetone. The later will make the cells permeable, in addition to acting as a fixative. The first two will act as fixative only. Good luck with our research.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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