Anti-cIAP2抗体(ab113237)

Please notice, these protocols may be optimised, and could not work with your specific tissue.

Ab32059: Steam slides in 0.01M Sodium Citrate Buffer, pH 6.0 at 125°C for 30 seconds followed by 10 seconds at 90° C into a decloaking chamber / pressure cooker. Removes slides and rinse in TBST before proceeding to the staining step.

Ab113237: Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes. Remove from heat and let stand at room temperature in buffer - 20 minutes. Rinse in 1X TBS with Tween (TBST) – 1 minute at RT.

Thanks for sending the image again. I'm sorry to see that the results are so poor with this replacement antibody. Do any of the bands on the blot correspond to the expected band size of cIAP2? How much lysate do you load into each well? The testing laboratory uses 5% non-fat dry milk instead of BSA to block the nitrocellulose membrane, and some antibodies are sensitive to blocking agent so it might be worth a try. Do you also dilute the primary and secondary antibodies in solution containing milk or BSA? This can be removed if so. Have you tried blotting for a loading control protein like beta actin on the same membrane, to check that the HRP and ECL kit are working well? We do guarantee this replacement antibody to work as well, so if the results do not improve I would be happy to issue a credit or refund for your original purchase. Please let me know if you have any questions or if there is anything else that we can do for you.