Does somebody know that the expression of STAT5 in rheumatoid arthritis synovial fibroblasts (RASFs)? Although some report indicated that the expression of STAT5 in RASFs, we can not detect any signals of STAT5 in RASFs using Western blot.
Please give me your opinions.
If there is expression of STAT-5 in RASFs and you do not get any signal there are quite a few possibilities for this:
1. Technical Issues: Are you loading enough protein in your gel? What is the affinity of your antibody for your protein of interest. Are you isolating you cell lysates in the proper lysis buffer?
2. Which STAT-5 (A/B) are you planning to look at?
3. You may also want to refer to link below (Figure 6). They have shown to have some success with phospho-STAT-5. However, I find it peculiar they did not normalize to total STAT-5 in the figure. Nonetheless, they did however show STAT-5 expression in normal SF (Figure 4) so you may want to order their antibody to test.
Article Induction of CCL13 Expression in Synovial Fibroblasts Highli...
Hi Jack,
Thank you for your opinions.
We always perform WB using T-cell lysates as positive control of STAT5. We can get signal of STAT5 in T-cell lysates, but not in RASFs.
We use polyclonal antibody for STAT5 which can bind both STAT5A and B. This antibody was purchased from CST.
We checked your recommended article. However, there are not so many study to investigate the expression of STAT5 in RASF.
Best wishes,
Kunihiko
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