We are working on an enzyme (145 KDa). Firstly we did Immunofluorescence staining on the paraffin embedded sections (4 µm thick). No protein was detected in PBS group while high level was detected in epithelial cells of treated group. Than we try WB and the protein was detected in both PBS and Treated group at the same level. What can be the possible reason?
It really depends. How did the results look like when you used a negative control for your immunofluorescence on both control and treated groups?
Also, how about your control in WB? How did that look?
Did you used a reference gene (e.g. β-actin, GAPDH) to normalize your samples? Did you run the samples of the two groups within a single SDS-PAGE?
Arman Rahimmi Thank You Arman. Yes I did use GAPDH as a reference gene and results are very clear. for GAPDH in all the samples. Yes I run a single SDS-PAGE for all the samples.
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