BD Biosciences
PE-CF594 Mouse Anti-Smad2 (pS465/pS467)/Smad3 (pS423/pS425)
562697
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We use this antibody to assess TGF-beta signaling in mouse NK cells. The antibody works well at the recommended dilution, giving a bright signal in stimulated cells.
Flow Cytometry
Mouse blood or splenocytes
1:20 dilution, 30 min at 4C
Fc block (Anti CD16/CD32)
None
Flow cytometer
In the attached image, mouse splenocytes were stimulated for 1 hour with 10ng/mL TGF-beta or 100ng/mL IL-15. Cells were immediately fixed using BD Fix-Lyse buffer, and stained for surface antigens to detect NK cells. Cells were then permeabilized using BD Perm Buffer III, and stained for pSMAD2/3. With TGF-beta stimulation, cells display robust SMAD2/3 phosphorylation, while IL-15 yields no response.
N/A
Bright signal, easy to use.
Nothing, I've never had any major problems.
Good antibody for pSMAD2/3