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Our Abpromise guarantee covers the use of ab3581 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 a concentration of 5 µg/ml.
Fixation: 4% paraformaldehyde for 15 minutes at room temperature. Permeabilisation: PBS containing 0.01% saponin for 15 mins Blocking: 1% BSA in PBS Primary: overnight at 4ºC
|EMSA||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration. PubMed: 20438742Sample Preparation: endogenous peroxidase blocked with 0.3% hydrogen peroxide. Antigen retrieval: Heat induced antigen retrieval performed in 0.01M Tris-EDTA pH 9.0. Primary antibody: overnight at 4°C|
|Flow Cyt||Use at an assay dependent concentration. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.|
|IP||Use at an assay dependent concentration. Sample preparation: RIPA buffer containing protease inhibitors was added to cell lysate. Primary antibody: 1:10 dilution for 2 h on ice Prewashed magnetic beads added to lysate mixture and incubated overnight at +4ºC. After washing, sample reducing buffer was added and boiled (5 min, 95ºC)|
|WB||1/500. Detects a band of approximately 90 kDa (predicted molecular weight: 83 kDa).Can be blocked with Human Glucocorticoid Receptor beta peptide (ab39765). Sample preparation: Lysates boiled in reducing buffer for 5 min. 20µg of protein loaded Blocking agent: 5% non-fat milk in Tris buffered saline (TBS) Primary antibody: overnight at 4°C|