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Our Abpromise guarantee covers the use of ab85615 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 26076134|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 62 kDa (predicted molecular weight: 46 kDa).|
|IHC-P||Use a concentration of 5 µg/ml.|
|ICC/IF||Use a concentration of 5 µg/ml.|
IHC image of 5HT1A Receptor staining in Human normal liver 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 ab85615, 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.
Immunohistochemistry (PFA perfusion fixed frozen sections) analysis of rat hippocampus labelling 5HT1A with ab85615 (red). For immunofluorescence analysis, the rats were anaesthetized using chloral hydrate and then were perfused with 150 mL of saline solution and 150–200 mL of 4% paraformaldehyde in 0.1 M phosphate buffer (PB). Then, the brains of the rats were isolated and fixed in 4% paraformaldehyde for 2–3 h. The rat brain sections were cut on a freezing microtome at a thickness of 16 μm and were washed with phosphate-buffered saline (PBS) and pre-incubated in 5% horse serum. Subsequently, the sections were incubated with Anti-5HT1A Receptor antibody (ab85615) for 24 h at 4°C, washed with PBS and incubated with the secondary antibody and neuro tracer for 2 h at room temperature while protected from light. The sections were examined on a Nikon C2 plus confocal laser-scanning microscope.