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 83 kDa (predicted molecular weight: 83 kDa).
Use at 2-5 µg/mg of lysate.
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
CSTF3 (cleavage stimulation factor subunit 3) is required for polyadenylation and 3'-end cleavage of mammalian pre-mRNAs and is part of the CSTF complex. The CSTF complex is composed of CSTF1 (50 kDa subunit), CSTF2 (64 kDa subunit) and CSTF3 (77 kDa subunit). CSTF3 functions as a homodimer and interacts directly with CSTF1 and CSTF2.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling CSTF3 with ab72299 at 1/200 (1µg/ml). Detection: DAB.
Western blot - CSTF3 antibody (ab72299)
All lanes : Anti-CSTF3 antibody (ab72299) at 0.04 µg/ml
Lane 1 : Whole cell lysate from HeLa cells at 50 µg Lane 2 : Whole cell lysate from HeLa cells at 15 µg Lane 3 : Whole cell lysate from HeLa cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg
Predicted band size: 83 kDa Observed band size: 83 kDa Additional bands at: 150 kDa, 31 kDa, 50 kDa, 60 kDa. We are unsure as to the identity of these extra bands.
Immunoprecipitation - CSTF3 antibody (ab72299)
Detection of Human CSTF3 by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), using ab72299 at 3 µg/mg lysate for IP and at 1 µg/ml for subsequent Western blot detection.