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Thanks so much for helping replace another MT (ab36882) antibody for our experiments. I recently have tried this replaced antibody to do some ICC (Immunocytochemistry) study; however, the result doesn't look very well. The signal is very very week (almost dark). I've tried two different concentration (1:500 and 1:1000) of this primary antibody on mouse Astroglia cells, but the data still doesn't look good. The detailed protocol please see below.
1. Cells were fixed with 4% PFA (15 min), then washed with PBS X 2 times
2. Blocking with 10% goat serum in PBST for 1 hr
3. Incubation primary Ab (ab12228 - 1:500 and 1:1000) in PBS-Tween (0.3%), overnight at 4 degrees.
4. Then wash with PBS x2, incubate with secondary Ab (Dylight-Green)- goat anti mouse 2nd Ab (diltuion 1:2000) for 3 hr.
The image still look very bad. Almost cannot see any signal. I'm wondering if we can request to replace another vial or another alternative one? If possible, please let me know. I can also send the image to you if necessary.Many thanks,
Asked on Nov 16 2012
Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.
I have read the details you have kindly provided and will be very happy to investigate the antibody further.
- Please fill the attached questionnaire and provide images?
Thank you very much for your cooperation. I will look forward to hearing from you soon.
Answered on Nov 16 2012