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My name is Johnny Huang and I’m from the University of Queensland, Australia. I purchased your antibodies against TIM-1 (ab66062 and 47634) recently. I tried the antibodies for western blotting but I couldn’t get single band. In fact, TIM-1 is a glycoprotein, and if you run a SDS-PAGE with purified TIM-1 protein, you will get a smear view on the PAGE. I noticed you have an example picture on your website, w+hich shows one single band on western blotting. https://www.abcam.com/TIM-1-antibody-ab66062.html Would you please let me know the conditions you use for the preparation of tissue lysis and western blotting in detail? Did you perform a de-glycosylation step before the western blotting? Thank you very much! Regards,
Asked on Sep 15 2011
Thank you for contacting us. We find that specific protocol is not necessary for this product. So I have attached the protocol that we generally recommend to our customers. We have used reduced denaturing conditions in WB without deglycosylation step. I can further help you with troubleshooting if you can provide more details about the protocol. Please find attached the questionnaire that we have created to get more information about the protocol customer use. Please fill in the form and send it to me via email asap. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Answered on Sep 15 2011