Overview

  • Product name
  • Description
    Rabbit polyclonal to TRPM8
  • Host species
    Rabbit
  • Specificity
    Reacts with residues 278-292 and the C terminus sequence of the human trpm8 protein.
  • Tested applications
    Suitable for: ELISA, IP, IHC-P, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Guinea pig, Human
  • Immunogen

    Synthetic peptide corresponding to Human TRPM8.
    Sequence:

    RNQLEKYISERTIQD Plus NDLKGLLKEIANKIK

  • Positive control
    • WB: Transfected COS cell lysate; human liver tissue lysate; human hepatocyte lysate. PMID 17510704, prostate cells used as positive control. PC-3 suitable negative control.

Properties

Applications

Our Abpromise guarantee covers the use of ab3243 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
ELISA 1/100 - 1/2000.
IP Use at an assay dependent concentration.

used at 6 ug/mg cell lysate.

IHC-P 1/10 - 1/2000.
WB 1/500 - 1/1000. Predicted molecular weight: 128 kDa.
IHC-Fr 1/100. See Abreview.

Target

  • Function
    Receptor-activated non-selective cation channel involved in detection of sensations such as coolness, by being activated by cold temperature below 25 degrees Celsius. Activated by icilin, eucalyptol, menthol, cold and modulation of intracellular pH. Involved in menthol sensation. Permeable for monovalent cations sodium, potassium, and cesium and divalent cation calcium. Temperature sensing is tightly linked to voltage-dependent gating. Activated upon depolarization, changes in temperature resulting in graded shifts of its voltage-dependent activation curves. The chemical agonists menthol functions as a gating modifier, shifting activation curves towards physiological membrane potentials. Temperature sensitivity arises from a tenfold difference in the activation energies associated with voltage-dependent opening and closing.
  • Tissue specificity
    Expressed in prostate. Also expressed in most in prostate tumors. Also expressed in non-prostatic primary tumors such as colon, lung, breast and skin tumors.
  • Sequence similarities
    Belongs to the transient receptor (TC 1.A.4) family. LTrpC subfamily. TRPM8 sub-subfamily.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Long transient receptor potential channel 6 antibody
    • LTrpC-6 antibody
    • LTrpC6 antibody
    • MGC2849 antibody
    • Short form of the TRPM8 cationic channel antibody
    • Transient receptor potential cation channel subfamily M member 8 antibody
    • Transient receptor potential p8 antibody
    • transient receptor potential-p8 antibody
    • Trp p8 antibody
    • Trp-p8 antibody
    • Trpm8 antibody
    • TRPM8_HUMAN antibody
    • TRPP8 antibody
    see all

Images

  • Anti-TRPM8 antibody (ab3243) at 1/750 dilution + TRPM8 transfected Cos7 cell lysate

    Predicted band size: 128 kDa

  • ab3243  staining TRPM8 in human prostate cancer tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections).

  • All lanes : Anti-TRPM8 antibody (ab3243) at 1000 cells

    Lane 1 : Human liver tissue lysate
    Lane 2 : Human hepatocyte lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit HRP at 1/1000 dilution

    Predicted band size: 128 kDa

    See Abreview

  • ab3243 staining TRPM8 in murine skin tissue by Immunohistochemistry (Frozen sections).
    Tissue was fixed in methanol, permeabilized with 0.4% Triton X-100, blocked using 10% serum for 30 minutes at 25°C and then incubated with ab3243 at a 1/100 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 594 conjugated goat anti-rabbit polyclonal used at a 1/500 dilution.

    See Abreview

References

This product has been referenced in:
  • Liu T  et al. RNA interference-mediated depletion of TRPM8 enhances the efficacy of epirubicin chemotherapy in prostate cancer LNCaP and PC3 cells. Oncol Lett 15:4129-4136 (2018). Read more (PubMed: 29541177) »
  • Klumpp D  et al. TRPM8 is required for survival and radioresistance of glioblastoma cells. Oncotarget 8:95896-95913 (2017). Read more (PubMed: 29221175) »
See all 17 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
human hepatocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted May 29 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Immuno-precipitation step
Protein A
Sample
Human Cell lysate - whole cell (Hek cells transfected with human TRPM8)
Specification
Hek cells transfected with human TRPM8
Total protein in input
800 µg

Miss. Larisa Maier

Verified customer

Submitted Sep 17 2013

Answer

Gracias por contactarnos, le pido disculpas por la espera.

Adjunto al email la datasheet del producto descatalogado ab3243. Al no estar disponible no tengo acceso a la datasheet en formato pdf, por lo que la envío copiada en un documento Word, siento los inconvenientes.

También le envío el enlace a la búsqueda llevada a cabo de anticuerpos anti TRPM8 humano en WB y IHC (Fr/P).

https://www.abcam.com/index.html?pageconfig=searchresults&search=TRPM8&pt=1&sk=app&sv=74&sn=IHC-P&l=4&fViewMore=1

Si necesitara más información acerca de cualquiera de los productos identificados, un presupuesto o tuviera cualquier otra consulta, por favor, no dude en volvernos a contactar.

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Question
Answer

Thank you for contacting Abcam. Please find the protocol used to test ab3243 in IHC below. If there is anything else I can help you with, please let me know.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (skin, HR-1)
Specification
skin, HR-1
Fixative
Methanol
Permeabilization
Yes - 0.4% TritonX100
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 20 2011

Answer

Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab3243 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. I can confirm that our laboratory tested this antibody with transfected Cos cell lysate in western blotting and managed to obtain a band at around 130kDa. Unfortunately, I am unable to retrieve any further protocol information (amount of protein loaded, etc.) and therefore, the image is not suitable for use on our datasheet. I have attached a copy of the image for your reference only. I am unsure of the exact protocols used by Yang et al 2006, but please rest assured that we guarantee our products to perform well in species and applications as stated on the datasheet. If you would like some technical support, I have looked through the protocols you used and have a few questions and suggestions that might help you resolve the problem. In most cases, the cause of multiple bands is because the primary and/or secondary antibody concentration is too high and causes non-specific binding. What was the dilution that you used? The dilution that we have on the datasheet is a recommended starting dilution and customers are encouraged to determine the optimal concentration/dilution. You can try decreasing the primary (1:2000 ~ 1:5000) and secondary antibody concentration or run a no-primary control (without the primary antibody). Running a no-primary control will be able to eliminate the possibility that your secondary antibody is binding non-specifically. Another possible cause of non-specific bands is that the amount of protein is too much. We normally recommend using 20-40 micrograms per well. Please try this amount if you have not already done so. What are the band sizes you obtained? The bands located above the expected molecular weight band could be multimers. Please try boiling your protein in SDS-PAGE for 10 minutes rather than 5 minutes to ensure proper disruption of multimers. Meanwhile, bands located below the expected molecular weight may indicate that your target protein has been digested. Please make sure that you incorporated sufficient protease inhibitors and have kept your samples on ice the whole time. Increasing the washing frequency and duration would also help eliminate non-specific bands. Can you confirm that you have used Tween in your washing solution and have washed at least 15min x 3 times? I hope the above recommendations may already help you, if you still experience problems please do not hesitate to contact me with the answers to the above questions and an image of the blot if possible. I would also appreciate if you could also provide details of your order (purchase order number, shipping address/purchasing agent info, contact information, etc.) so that I can immediately arrange for a replacement or refund to you. Also, please advice on how you would to proceed with this enquiry. I look forward to hearing from you again in order to resolve the matter.

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Answer

Thank you for your e-mail to Hugh who is away today. I'm very sorry to hear you are still experiencing lots of non specific bands with ab3243 even after the protocol modifications. I would be happy to offer you a replacement vial or refund if you purchased the antibody in the last 3 months. Can you please confirm your order details and what you would prefer? I look forward to hearing from you to arrange this,

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I have read your technical questionaire and I have a few comments. We have details of this antibody applied by IHC from customer feedback, but not western blotting. You are obtaining an alarming number of extraneous bands. I would like to recommend that in order to obtain more specific results that you use 3% BSA as a blocking agent. This is routinely employed by our lab team. I would also like to recommend that you increase the percentage of Tween 20 to 0.1% in an effort to address the non-specific nature of this antibody. I would also like to recommend that you perform an overnight block of your membrane using this agent. If this does not improve your results please get back in touch with me and I am happy to provide you with a replacement antibody or credit against your purchase provided that you purchased this antibody within the past 90 days.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (prostate cancer)
Specification
prostate cancer
Blocking step
Other as blocking agent for 30 minute(s) · Concentration: 1.2%

Dr. Gabriel Bidaux

Verified customer

Submitted Aug 21 2006

Answer

Thank you for your enquiry. I am sorry but we do not have the immunogen peptide for this antibody in our catalogue. It may have been exhausted. Yes, were you to have the two peptides used to raise this antibody synthesized I can see no reason why they would not serve as peptide inhibition controls for this antibody.

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1-10 of 19 Abreviews or Q&A

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

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