The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000 - 1/10000. Detects a band of approximately 77 kDa (predicted molecular weight: 77 kDa).
Use at 2-5 µg/mg of lysate.
Use at an assay dependent concentration.
Component of the cleavage and polyadenylation specificity factor (CPSF) complex that play 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. Has endonuclease activity, and functions as mRNA 3'-end-processing endonuclease. Also involved in the histone 3'-end pre-mRNA processing. U7 snRNP-dependent protein that induces both the 3'-endoribonucleolytic cleavage of histone pre-mRNAs and acts as a 5' to 3' exonuclease for degrading the subsequent downstream cleavage product (DCP) of mature histone mRNAs. Cleavage occurs after the 5'-ACCCA-3' sequence in the histone pre-mRNA leaving a 3'hydroxyl group on the upstream fragment containing the stem loop (SL) and 5' phosphate on the downstream cleavage product (DCP) starting with CU nucleotides. The U7-dependent 5' to 3' exonuclease activity is processive and degrades the DCP RNA substrate even after complete removal of the U7-binding site. Binds to the downstream cleavage product (DCP) of histone pre-mRNAs and the cleaved DCP RNA substrate in a U7 snRNP dependent manner.
Belongs to the metallo-beta-lactamase superfamily. RNA-metabolizing metallo-beta-lactamase-like family. CPSF3 subfamily.
Sumoylated on Lys-462, Lys-465 and Lys-545, preferentially by SUMO3.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma (left) and mouse teratoma (right) tissues labelling CPSF73 with ab72295 at 1/200 (1µg/ml). Detection: DAB.
Western blot - CPSF73 antibody (ab72295)
All lanes : Anti-CPSF73 antibody (ab72295) 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 : Mouse NIH3T3 whole cell
lysate at 50 µg
Developed using the ECL technique
Predicted band size : 77 kDa Observed band size : 77 kDa
Exposure time : 30 seconds
Immunoprecipitation - CPSF73 antibody (ab72295)
Detection of CPSF73 by Western Blot of Immunprecipitate.
Lane 1: ab72295, at 0.1µg/ml, staining CPSF73 in HeLa whole cell lysate immunoprecipitated using ab72225 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded).br>
Lane 2: Control IgG.
Detection: Chemiluminescence with an exposure time of 30 seconds.