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 95 kDa (predicted molecular weight: 95 kDa).
Use at 2-5 µg/mg of lysate.
Regulatory subunit of serine/threonine-protein phosphatase 4. May regulate the activity of PPP4C at centrosomal microtubule organizing centers. The PPP4C-PPP4R2-PPP4R3A PP4 complex specifically dephosphorylates H2AFX phosphorylated on 'Ser-140' (gamma-H2AFX) generated during DNA replication and required for DNA DSB repair.
Belongs to the SMEK family. Contains 1 WH1 domain.
Cytoplasm. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. Nucleus. In interphase localized in the cytoplasm and in the nucleus (with higher levels). During metaphase located in pericentriolar regions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma (left) and breast carcinoma (right) tissues labelling Smek1 with ab70635 at 1/200 (1µg/ml) and 1/80 (2.5µg/ml). Detection: DAB and DyLight® 594-conjugated goat anti-rabbit IgG (1/100).
Western blot - Smek1 antibody (ab70635)
All lanes : Anti-Smek1 antibody (ab70635) 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
Developed using the ECL technique.
Predicted band size: 95 kDa Observed band size: 95 kDa
Exposure time: 10 seconds
Immunoprecipitation - Smek1 antibody (ab70635)
Detection of Smek1 by Western Blot of Immunprecipitate.
ab70635 at 1µg/ml staining Smek1 in HeLa whole cell lysate immunoprecipitated using ab70635 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane).
Detection: Chemiluminescence with exposure time of 0.5 seconds.