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His-tagged Hamster Cyclin B1 expressed in bacteria, harvested from inclusion bodies, extracted with 6M guanidine HCl and purified on Nickel beads.
Our Abpromise guarantee covers the use of ab72 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|IHC (PFA fixed)||1/400.|
|Flow Cyt||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 48 kDa).
Abcam recommends using 5% Milk as the blocking agent.
|IHC-FoFr||Use a concentration of 1 - 2 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml.|
|IHC-Fr||Use a concentration of 1 - 2 µg/ml.|
IHC image of ab72 staining in Human Normal Tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab72, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Lanes 1-9 shows the Cyclin B1 wave as HeLa cells enter and exit mitosis. HeLa cells were synchronised in the early S phase by double thymidine block then released to synchronously enter mitosis. Time points were taken at the points indicated.
HeLa whole cell lysate loaded at 25ug per lane. Ab72 used at 1/500 in reducing conditions in conjunction with goat anti mouse (HRP) 1/20000.
This image is courtesy of an Abreview submitted on 12 September 2005. We do not have any further information relating to this image.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"