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Recombinant Human SIRT1
This antibody clone is manufactured by Abcam.
Western blot protocol advice:
For best results in Western blot using this antibody, we recommend the following:
Our Technical team (email@example.com) will be happy to provide further information and advice.
Our Abpromise guarantee covers the use of ab110304 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration.|
|In-Cell ELISA||Use a concentration of 4 µg/ml.|
|WB||Use a concentration of 1 - 0.125 µg/ml. Predicted molecular weight: 81 kDa.
Detects a band of approximately 110 kDa (110-121 kDa) which is likely to be due to post translational glycosylation. SIRT1 is known to bind to several other proteins, and the 121 kDa band could also be due to the presence of one of these complexes (ensure samples are adequately reduced and denatured).
|Flow Cyt||Use a concentration of 1 µg/ml.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use a concentration of 0.5 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml.|
IHC image of SIRT1 staining in Human normal colon 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 ab110304, 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
Blocked with 3% milk (TBS-tween) at 4C for 16 hours
Primary Antibody: 0.25 µg/mL in 10X Blocking Buffer (ab126587). 3hrs at room temperature.
Secondary Antibody: 1:5,000 in 10X Blocking Buffer (ab126587). 3hrs at room temperature.
ab110304 detects a band of approximately 110 kDa (110-121 kDa) which is likely to be due to post translational glycosylation. SIRT1 is known to bind to several other proteins, and the 121kDa band could also be due to the presence of one of these complexes (ensure samples are adequately reduced and denatured).
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 3% Milk before being incubated with ab110304 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"