Anti-Galectin 7 抗体 (ab10482)

Thank you for your enquiry regarding ab10482 and for providing some further information. I am very sorry to hear that you are having some problem with this antibody. I understand from your e-mail that you used lymfoblast cell lysate. Can you please specify the species? Blocking is very important step and the time and the temperature is crucial. Normally I suggest blocking the membrane at least for 1 hr at 4oC – it seems that 10 min is too short. For further information on blocking and Western blot, please visit our website at: https://www.abcam.com/index.html?pageconfig=resource&rid=10413 https://www.abcam.com/assets/pdf/protocols/WB-beginner.pdf Have you tried to use 5% BSA for blocking instead of milk? Milk sometimes can produce non-specific bands on the blot and in our lab we usually use BSA for blocking. I would also recommend testing the secondary antibody alone i.e. run a no primary control to see if the non-specific bands are due to the secondary antibody or not. I hope this will help. Should you still have problem with this antibody, then please do not hesitate to contact me again.

Thank you for your enquiry and your patience. Here we are forwarding the protocol for Preparation of Epidermal Lysates: Transgenic and nontransgenic mice were killed by cervical dislocation. The dorsal skins were treated with a depilatory agent (1 min) followed by washing. The skin was excised, and the epidermis was scraped off with a razor blade into lysis buffer [50 mm Tris (pH 7.4), 1% Triton X-100, 150 mM NaCl, 1 mM EDTA, 1 mM PMSF, 1 mM Na3VO4, 1 mM NaF, and 1 ug/ml each of aprotinin, leupeptin, and pepstatin]. The lysates were homogenized using a needle (18-gauge) and syringe and were centrifuged at 14,000 x g for 15 min at 4°C. The supernatant was used immediately for Western blot analysis. We hope this information will be useful for you. If you need anything further or any help then please let us know.