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Our Abpromise guarantee covers the use of ab20210 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.|
|IHC-Fr||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ICC||Use a concentration of 5 - 10 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
ab20210 stained HepG2 cells. The cells were 100% methanol fixed for 5 minutes at room temperature and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab20210 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150117 used at a 1/1000 dilution for 1hour at room temperature. The cells were counterstained with ab202277 (Rabbit monoclonal to alpha Tubulin (Alexa Fluor® 594) (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
IHC image of Cytokeratin 19 staining in human normal liver formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ 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 ab20210, 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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
ab20210 staining Cytokeratin 19 in human breast carcinoma tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with methanol and blocking with 10% serum for 5 minutes was performed. The sample was incubated with primary antibody (1/100) in PBS with 10% goat serum for 1 hour. A HRP-conjugated rabbit polyclonal to mouse IgG was used undiluted as secondary antibody.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"