ab193977 was shown to specifically react with Cyclin B1 in wild-type HAP1 cells as signal was lost in CCNB1 (Cyclin B1) knockout cells. Wild-type and CCNB1 (Cyclin B1) knockout samples were subjected to SDS-PAGE. Ab193977 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.
IHC image of Cyclin B1 staining in a section of formalin-fixed paraffin-embedded human colon adenocarcinoma*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab193977, at a working dilution of 1/100 overnight at +4°C. The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-Cyclin B1 antibody [Y106] (HRP) (ab193977)
All lanes : Anti-Cyclin B1 antibody [Y106] (HRP) (ab193977) at 1/5000 dilution
Lane 1 :HeLa whole cell lysate (ab150035) Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 58 kDa Observed band size: 58 kDa
Exposure time: 1 minute
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab193977 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.