Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
CPSF1 (Cleavage and polyadenylation specificity factor subunit 1) is a component of the cleavage and polyadenylation specificity factor (CPSF) complex. It plays a key role in pre-mRNA 3'-end formation, recognizing the AAUAAA signal sequence and interacting with poly(A) polymerase and other factors to bring about cleavage and poly(A) addition. This subunit is involved in the RNA recognition step of the polyadenylation reaction.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma (left) and mouse renal cell carcinoma (right) tissues labelling CPSF1 with ab81552 at 1/200 (1µg/ml). Detection: DAB.
Western blot - Anti-CPSF1 antibody (ab81552)
All lanes : Anti-CPSF1 antibody (ab81552) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg Lane 5 : NIH3T3 whole cell lysate at 50 µg
HeLa cell lysate (1mg) was immunoprecipitated using ab81552 at 3µg/mg lysate (lane 1) or IgG control (lane 2). 20% of the immunoprecipitate was analysed by Western blot using ab81552 at 1µg/ml. Bands were detected by chemiluminescence with an exposure time of 10 seconds.
IHC image of ab81552 staining in Human Breast cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab81552, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.