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Full length native Nestin protein purified from embryonic rat spinal cord.
Our Abpromise guarantee covers the use of ab6142 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 - 1/200. Cells fixed with 4% paraformaldehyde buffered with 50mM sodium borate at pH 9.5.|
|Electron Microscopy||Use a concentration of 0.1 - 1 µg/ml. Tissue fixed with 4% paraformaldehyde at pH 10.0 or 4% paraformaldehyde with 0.1% glutaraldehyde at pH 7.4.|
|Flow Cyt||Use at an assay dependent concentration. PubMed: 21092738|
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 18562299|
|IHC-P||Use a concentration of 0.1 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Tissue fixed with 4% paraformaldehyde at pH 7.4 for light microscopy.|
|IHC-Fr||Use a concentration of 0.1 - 1 µg/ml. Tissue fixed with 4% paraformaldehyde at pH 7.4 for light microscopy.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 200 kDa. Block with milk or BSA but do not dilute primary antibody in buffer containing milk.|
ab6142 at 1/500 dilution staining Nestin in rat brain tissue (green) by immunohistochemistry (frozen sections). Sections were methanol fixed, permeabilized in 0.1% Triton X-100 prior to blocking in 2.5% BSA for 16 hours at 4°C and then incubated with ab6142, for 1 hour at 37°C. Alexa fluor® 680 goat polyclonal to mouse Ig, diluted 1/1000 was used as the secondary antibody.
ab6142 staining Nestin in mouse brain tissue sections by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone, permeabilized with Triton X-100 in PBS and blocked with 5% BSA for 60 minutes at room temperature. Samples were incubated with primary antibody (1/1000 in PBST) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-mouse IgG monoclonal was used as the secondary antibody at a dilution of 1/200.
ab6142 staining adult mouse brain tissue section by Immunohistochemistry (Formalin/PFA-fixed, paraffin embedded sections). Tissue underwent fixation in paraformaldehyde, heat mediated antigen retrieval in Sodium Citrate, permeabilization in 1% Triton buffer and blocking in 10% serum for 1 hour at 25°C. The primary antibody, diluted 1/200 (PBS, 2% Donkey serum, 0.2% Triton) for 16 hours at 4°C. An Alexa Fluor® 488 conjugated donkey polyclonal to mouse Ig, diluted 1/500 was used as the secondary.
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