Anti-TRPM8 抗体 (ab3243)

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.

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.

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.

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,

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.