The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
1/2000 - 1/10000. Detects a band of approximately 97 kDa (predicted molecular weight: 97 kDa).
Use at 2-5 µg/mg of lysate.
SMEK2 contains a WH1 domain. There are five named isoforms produced by alternative splicing. Mapping of a diet-induced hypercholesterolemia locus (Dihc2) in rats identified SMEK2 as a strong candidate for responsiveness to dietary cholesterol.
Cytoplasm. Cytoskeleton. Centrosome. Nucleus. Note: In interphase localized in the cytoplasm and (with higher levels) the nucleus. During metaphase located in pericentriolar regions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling SMEK2 with ab70622 at 1/200 (1µg/ml). Detection: DAB.
Western blot - SMEK2 antibody (ab70622)
All lanes : Anti-SMEK2 antibody (ab70622) 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
Predicted band size: 97 kDa Observed band size: 97 kDa
Immunoprecipitation - SMEK2 antibody (ab70622)
Immunoprecipitation of HeLa whole cell lysate (1 mg) using ab70622 at 3 µg/ml (lane 1) or control IgG (lane 2). 20% of immunoiprecipitate was analysed by Western blot using ab70622 at 1 µg/ml.