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PGP 9.5 from human brain.
This antibody clone is manufactured by Abcam.
This antibody labels the neuronal cell bodies and axons in central and peripheral neural system. Small nerve fibers in peripheral tissues, neuroendocrine cells in normal pituitary thyroid, pancreas, and gastrointestinal tract, as well as derived tumors are also stained with this antibody.
Our Abpromise guarantee covers the use of ab8189 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/20 - 1/50.|
|IHC-FoFr||1/10 - 1/20.|
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 0.5 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 5 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: PGP9.5 knockout HAP1 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab8189 observed at 25 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab8189 was shown to specifically react with PGP9.5 in wild-type HAP1 cells as signal was lost in PGP9.5 knockout cells. Wild-type and PGP9.5 knockout samples were subjected to SDS-PAGE. ab8189 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 5 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 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.
IHC image of PGP9.5 staining in Rat pancreas 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 ab8189, 0.02µg/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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"