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Fusion protein corresponding to Cow Cyclin A2.
Our Abpromise guarantee covers the use of ab38 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 5 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 60 kDa).|
|IP||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration. PubMed: 22802651|
IHC image of Cyclin A 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 ab38, 10µg/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.
Immunoprecipitation using ab38 at 4µg/mg whole cell lysate diluted in RIPA buffer and incubated with Protein G matrix at 4°C for 12 hours.
This antibody can immunoprecipitate Cyclin A from both Human (293T) and Rat (PC12) cell extract. The species assigned as Cyclin A runs at ~55 kDa.
Lanes 1-3 = 293T
Lanes 4-6 = PC12
Lane 1: Cyclin A antibody IP
Lane 2: Control antibody IP
Lane 3: 293T whole cell lysate at 10%
Lane 4: Cyclin A antibody IP
Lane 5: Control antibody IP
Lane 6: PC12 whole cell lysate at 10%
The Western Blot antibody was ab38 at 2µg/ml
This image is courtesy of an anonymous Abreview
The membrane was blocked with 5% milk for 1 hour at 21°C prior to incubation with the primary antibody for 12 hours at 4°C.
This image is courtesy of Julian Gannon, CRUK.Western Blot of Swiss 3T3 cells showing presence of cyclin A in rapidly dividing cells (1) but absent in quiesent cells (2). Recombinant cyclin A (3). Molecular weight markers 114, 98, 68, 57, 43 and 32 kD. Western Blot of Swiss 3T3 cells showing presence of cyclin A in rapidly dividing cells (1) but absent in quiesent cells (2). Recombinant cyclin A (3). Molecular weight markers 114, 98, 68, 57, 43 and 32 kD.