Santa Cruz Biotechnology
Waf1/Cip1/CDKN1A p21 Antibody (F-5)
sc-6246
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DNA damage induced by gemcitabine or doxorubicin in bEnd.3 cells was assessed using p21 staining. The same antibody was also evaluated in mouse tissue samples, where target detection proved highly challenging.
Immunofluorescence
bEnd.3 cells, fixed in PFA 4% (10 min @ RT)
1 % Triton (10 min @ RT), followed by incubation with primary antibody for 1h @ RT (dilution 1:50 in BSA)
1% BSA
AlexaFluor 448 donkey anti-mouse IgG (H+L), dilution 1:800
none
Fluorescent microscopy
Successful detection of p21 in bEnd.3 cells following doxorubicin or gemcitabine exposure confirms the induction of DNA damage. However, staining intensity was relatively modest and should always be interpreted in comparison with untreated control cells to distinguish specific signal from background.
Efficient
Modest intensity levels. Use of a high dilution. Difficult to detect in IHC.
We evaluated several p21 antibodies from Santa Cruz Biotechnology, and this antibody demonstrated the best overall performance.