Anti-TRPV1 antibody (ab10296)

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Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-TRPV1 antibody (ab10296)

    Key features and details

    • Rabbit polyclonal to TRPV1
    • Suitable for: IHC-FoFr
    • Reacts with: Rat
    • Isotype: IgG

    Overview

    • Product name

      Anti-TRPV1 antibody
      See all TRPV1 primary antibodies

    • Description

      Rabbit polyclonal to TRPV1

    • Host species

      Rabbit

    • Specificity

      This antibody has been optimised on rat tissue. We recommend ab31895 for detection of TRPV1 in mouse tissue.

    • Tested Applications & Species

      Application Species
      IHC-FoFr
      Rat
      See all applications and species data

    • Immunogen

      Synthetic peptide within Rat TRPV1 aa 1-100 (N terminal). The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements.
      Database link: O35433

      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

    • 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
    • Concentration information loading...

    • 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

    The Abpromise guarantee

    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.

    Guaranteed

    Tested applications are guaranteed to work and covered by our Abpromise guarantee.

    Predicted

    Predicted to work for this combination of applications and species but not guaranteed.

    Incompatible

    Does not work for this combination of applications and species.

    Application Species
    IHC-FoFr
    Rat
    All applications
    Mouse
    Non human primates
    Application Abreviews Notes
    IHC-FoFr
    1/1000.

    (see Guo et al reference).
    Notes
    IHC-FoFr
    1/1000.

    (see Guo et al reference).

    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

      • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-TRPV1 antibody (ab10296)
        Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-TRPV1 antibody (ab10296)

        TRPV1 staining in the rat dorsal root ganglion using ab10296.

      References (6)

      Publishing research using ab10296? Please let us know so that we can cite the reference in this datasheet.

      ab10296 has been referenced in 6 publications.

      • Zhu TH  et al. Mast cell stabilizer ketotifen reduces hyperalgesia in a rodent model of surgically induced endometriosis. J Pain Res 12:1359-1369 (2019). PubMed: 31118754
      • Zhang J  et al. Upregulation of TRPV1 in spinal dorsal root ganglion by activating NGF-TrkA pathway contributes to pelvic organ cross-sensitisation in rats with experimental autoimmune prostatitis. Andrologia 51:e13302 (2019). PubMed: 31074030
      • 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 . 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 . PubMed: 19523146
      • Chen W  et al. Immortalization and characterization of a nociceptive dorsal root ganglion sensory neuronal line. J Peripher Nerv Syst 12:121-30 (2007). PubMed: 17565537
      • Naeini RS  et al. An N-terminal variant of Trpv1 channel is required for osmosensory transduction. Nat Neurosci 9:93-8 (2006). PubMed: 16327782

      Customer reviews and Q&As

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      1-4 of 4 Abreviews or Q&A

      Question

      BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Non-specific band at 50 kDa. SAMPLE Rat and horse dorsal root ganglion extract using Tri Reagent from Molecular Research Center. Band was in horse extract. There were no band s in the rat extracts. PRIMARY ANTIBODY Abcam ab10296, diluted 1:1000 in blocking buffer, incubated with shaking 1 hr at RT, washed with PBS/tween 20 4X. DETECTION METHOD ECL from Amersham POSITIVE AND NEGATIVE CONTROLS USED Used rat dorsal root ganglion tissue extract. Can you recommend a positive control from your catalog? ANTIBODY STORAGE CONDITIONS -20C SAMPLE PREPARATION Followed Tri REagent protocol, solubilizing with 1%SDS with tributylphosphine. Then, added loading buffer containing SDS and 2-mercaptoethanol. Heated at 100C for 4 min. AMOUNT OF PROTEIN LOADED 8 ug ELECTROPHORESIS/GEL CONDITIONS reducing 10% gel TRANSFER AND BLOCKING CONDITIONS Tris/glycine/methanol transfer buffer. Transfered 90 min @ 90 V. Blocked with 5% milk in PBS/tween 20. SECONDARY ANTIBODY Affinity Bioreagents donkey anti-rabbit HRP, diluted in blocking buffer 1:2500. Incubated and wash as with primary antibody. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

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      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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      Question

      We have a customer who has used this antibody for a while for western blot and IHC on paraffin sections. Previous batches have worked well for both, but the most recent lot (160975) has not been working as well (>50% less in western and no signal now in paraffin sections). Do you have any other lots available and if so could they be switched out for the lot that seems to be weaker? Thank you.

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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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      Question

      I have tried to detect the Vanilloid Receptor 1 in Western blots of our DRG neuron extracts using antibodies from a number of companies, but the results have been very discouraging, i.e, numerous bands in the vicinity of 100 kDa. Could you provide any information or show me any results that might persuade us to try your antibody. Would you consider supplying us with a minute quantitiy of the antibody (enough for one blot) to try on our extracts prior to our spending $300 for your product?

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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

      Are ab3486 and ab10296 suitable for ICC formalin fixed?

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      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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