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Two synthetic peptides corresponding to two regions of the Tau gene product corresponding to sequences shared between the mouse (P01637, NCBI) and human (NP_05819, NCBI) proteins, and not containing any of the serine and threonine residues known to undergo phosphorylation.
Two different affinity purified anti peptide antibodies were combined to make this product. The concentrations of both of these antibodies was 100 µg/ml, making the total antibody concentration 200 µg/ml.
Our Abpromise guarantee covers the use of ab75714 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/20000 - 1/50000. Predicted molecular weight: 79 kDa.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC||1/1000 - 1/2000.|
Immunohistochemistry (10% Formalin-fixed paraffin-embedded sections) analysis of Human Alzheimer's disease brain (CA1 region of hippocampus) tissue labelling Tau (brown) with ab75714.
Immunofluorescence analysis of primary entorhinal cortex neurons prepared from five-day-old rats, staining Tau with ab75714 at 1/1500 dilution.
Cells were fixed with paraformaldehyde, blocked in PBS + 5% fetal calf serum (ab7479), then permeabilized in PBS containing 0.2% Triton X-100. Samples were incubated with primary antibody for 2 hours at room temperature. A Cy5®-conjugated anti-chicken IgG was used as the secondary antibody.
IHC image of Tau staining in Human Hippocampus (Alzheimer's) (ab4583) 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 ab75714, 5 µg/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.