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This antibody was selected after immunization with a fusion protein containing the E. coli maltose binding protein and a fragment of the human MRP4 protein corresponding to amino acids 372-431.
Our Abpromise guarantee covers the use of ab15602 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 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 22272278PFA fixed cells|
|WB||1/20 - 1/50. Predicted molecular weight: 159 kDa.|
|IHC-Fr||1/20. Fix with acetone.|
|ICC||1/20 - 1/50. (acetone fixed cytospin preparations)|
Lanes 1 - 2: Merged signal (red and green). Green - ab15602 observed at 200-250 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab15602 was shown to specifically react with MRP4 in wild-type HAP1 cells as signal was lost in ABCC4 (MRP4) knockout cells. Wild-type and ABCC4 (MRP4) knockout samples were subjected to SDS-PAGE. Ab15602 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 20 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rat IgG H&L (IRDye® 800CW) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
HC image of MRP4 staining in Human lung adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. 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 ab15602, 10µg/ml, for 15 mins at room temperature. A Goat anti-Rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"