The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at 2-5 µg/mg of lysate.
1/2000 - 1/10000. Detects a band of approximately 65 kDa (predicted molecular weight: 59 kDa).
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Nuclear receptor coactivator NCoA62 (Nuclear Protein SkiP, SKIIP , ski-interacting protein) is a member of the SNW gene family, encodes a coactivator that enhances transcription from some Pol II promoters. This coactivator can bind to the ligand-binding domain of the vitamin D receptor and to retinoid receptors to enhance vitamin D-, retinoic acid-, estrogen-, and glucocorticoid-mediated gene expression. It can also interact with poly(A)-binding protein 2 to directly control the expression of muscle-specific genes at the transcriptional level. Finally, the protein may be involved in oncogenesis since it interacts with a region of SKI oncoproteins that is required for transforming activity.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma (left) and mouse teratoma (right) tissues labelling NCOA62 with ab70828 at 1/200 (1µg/ml). Detection: DAB.
Western blot - Anti-NCOA62 antibody (ab70828)
All lanes : Anti-NCOA62 antibody (ab70828) 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
Detection of Human NCOA62 by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), using ab70828 at 3 µg/mg lysate (Lane 1). Lane 2 represents IgG IP control. Subsequent Western blot detection of NCOA62 was performed using ab70828 at 1 µg/ml.