Hi
I did a western blotting in endometrial cancer cell lines for Cyclin D1 (1:1000, incubated over night, Santa Cruz) and used mouse as a secondary antibody (1:10,000, incubated for 4 hours). The results were negative. Wanted to to know if i could modify the steps. If anybody has had a positive results in Cyclin D1 please let me know.
Thank you for your help
Hi, I don't known the code of your santacruz antibody, is there an image in the datasheet ? Assuming that the antibody works well, you can try to change the primary concentration, try 1:500 or less.
In general the santacruz datasheet recommend a starting dilution but also a dilution range, that you have to check, regarding your samples
thks for replying. Ya there is an image in the data sheet. the code is (A-12) sc-8396.. Ok i will try lesser concentration
Hi, on Santacruz site I find this (https://www.scbt.com/scbt/it/product/cyclin-d1-antibody-a-12#citations) reviews about the antibody, they used the 1:500 or 1:200 dilution, I think with your samples and with 40ug you have to see the signal.
I do not recommend to lesser concentration of secondary, if you are sure that it is good with other primary antibodies
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