Overview

  • Product name
  • Description
    Guinea pig polyclonal to VR1
  • Host species
    Guinea pig
  • Tested applications
    Suitable for: IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat
    Predicted to work with: Guinea pig
  • Immunogen

    Synthetic peptide:

    YTGSLKPEDAEVFKDSMVPGEK

    , corresponding to C terminal amino acids 817-838 of Rat Vanilloid Receptor 1.

  • Positive control
    • This antibody gave a positive result when used in the following methanol fixed cell lines: PC12
  • 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

Associated products

Applications

Our Abpromise guarantee covers the use of ab10295 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
IHC-Fr 1/1000.
ICC/IF 1/100.

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

  • ab10295 stained PC12 cells. The cells were 100% Methanol fixed for 5 minutes and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab10295 at 1in100 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab96959 Dylight 488 goat anti Guinea pig used at a 1/250 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • ab10295, at a dilution of 1/1000, staining VR1 in rat dorsal root ganglion by Immunohistochemistry (PFA perfusion fixed frozen sections).

References

This product has been referenced in:
See all 5 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer



I am happy to confirm that the blocking peptide for ab10295 is now available under the product number ab134720:

https://www.abcam.com/index.html?datasheet=134720 (or use the following: https://www.abcam.com/index.html?datasheet=134720).

I would like to encourage you to submit an Abreview. We and our customer appreciate information about our products very much and you would earn Abpoints that can be redeemed as discount on future orders or Amazon gift vouchers.

We are looking forward to your order and wish you good luck with your experiments.

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Answer

Thank you for your reply.

I would be happy to arrange for a different lot to be sent to you. You do not need to place a new order, I can arrange it all for you. If you could just confirm that the following address is correct:
xxxxx


I look forward to receiving your reply.

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Question

1) Abcam product code ab3002101000 (lot GR64830-N/A)
2) Abcam order reference number or product batch number- 980870
3) Description of the problem - Absence of staining
4) Sample preparation:
Species: Rat and mouse
Type of sample: Fresh frozen sections (yes), perfusion fixed frozen sections (yes), PFA/formalin fixed paraffin embedded sections(yes), cells in culture, other:
Sample preparation
Positive control: DRG; Spinal Cord
Negative control: Tissue from VR1- KO mice
5) Fixation step
Yes/No - Yes
If yes: Fixative agent and concentration - PFA; formalin; tamponade formol;
Fixation time - 2h or overnight for tamponade formol
Fixation temperature - 4ºC
6) Antigen retrieval method: Since not used to warming (water bath - 80ºC - 30 min)
7) Permeabilization method:
Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
Permeabilizing agent and concentration: Tween - 100
8) Blocking agent (eg BSA, serum…): NDS
Concentration - 2% or 10%
Blocking time- 2h
Blocking temperature - room temperature
9) Endogenous peroxidases blocked? Yes
Endogenous biotins blocked? No
10) Primary antibody (If more than one was used, describe in “additional notes”) :
Concentration or dilution: 1: 100; 1:250;1:500;1:1000;1:2000
Diluent buffer: Blocking solution
Incubation time 24h, 48h or 72h
11) Secondary antibody:
Species:
Reacts against: anti-guinea pig
Concentration or dilution: 1:200
Diluent buffer: Blocking solution
ncubation time: 2h
Fluorochrome or enzyme conjugate: biotinilated or alexa fluor 488
12) Washing after primary and secondary antibodies:
Buffer - PBST
Number of washes - 3
13) Detection method: DAB and fluorescence microscopy
14) How many times have you run this staining?
Do you obtain the same results every time?Yes
What steps have you altered to try and optimize the use of this antibody? Concentration of primary, time and type of fixation; concentration of blocking solution; secondary antibody; time of incubation with primary; antigen retrieval (using or not);
Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.
Well about the choosing of this antibody: it is related with the fact that was already used and validated in studies of my area of interest - RVM in a papper referenced in your site.
With the best regards

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Answer

Thank you very much for providing that extra information. It has helped greatly in understanding the problems encountered and the optimisation you have already performed.

The anti-VR1 antibody (ab10295) has only been validated in frozen sections with rat tissue. This does not mean that it will not work in order tissue preparations or species but we do not currently have any information to support this. It may be that although the immunogen used to raise the antibody,dispite sharing 95% homology with the mouse protein, is not able to detect this species. Additionally, as paraffin embedded sections have not been used this may not be ideal with this antibody, as is the case with the Western blotting too. May I ask, what was the result of the Western blot? Could you send me an image of the blot?

We would expect the antibody to be able to detect the protein in your rat tissue with frozen sections, which you have not observed. I am contacting the source of this antibody to find out the exact protocol used to prepare the tissue in order to see if this could be contributing to the lack of signal.

In the meantime, if you would like, I can offer you a replacement vial of a different lot or a different VR1 antibody altogether such as rabbit polyclonal ab31895. This has been used in both mouse and rat tissue with paraffin embedded and frozen sections.

I look forward to receiving your reply.

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Answer

Thank you for contacting us and reporting the problems you have encountered in using the anti-VR1 antibody (ab10295). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I appreciate the time you have spent in the lab trying to optimise the results with this antibody. However, in order to more fully understand what may be contributing to the problems I would really appreciate it if you could give me a few more details on how the antibody has been used. To this end I have attached a questionnaire to this email. OnceI receive the completed questionnaire,I will look at the protocol and see if there are any suggestionsI can make that may improve the results.

This information will also allow us to investigate this case internally and initiate anyadditional testing where necessary. Whilst the product was purchased outside of the last 6 months and is therefore not covered by the Abpromise guarantee, I understand your concerns and the effort you have made and I would therefore be willing to extend this cover if the antibody has been usedaccording to our Abpromise, andwould be happy to replace or refund the antibody.

May I ask,is there a specificreason whyab10295(guinea pig polyclonal) was chosen and not one of our other antibodies againstVR1?

I look forward to receiving your reply.

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Answer

DISCOUNT CODE: ****** Expiration date: ***** I am very pleased to hear you would like to accept our offer and test ab10295 in western blot. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for western blot and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.  

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Answer

Thank you for contacting Abcam about ab10295. We do not sell sample sizes of our antibodies but since ab10295  has not been tested in western blot, I can offer a discount off a future purchase if you buy ab10295 now, test it in western blot and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab10295 in western blotI will then send a discount code. This code must be issued before purchasing ab10295 so please wait for my reply before ordering. 2. Purchase ab10295 either by phone, fax, or online (www.abcam.com). 3. Test it in western blot. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab10295 turns out to be unsuitable for western blot you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.  

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (brain)
Specification
brain
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10

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Verified customer

Submitted Feb 13 2008

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