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Full length native protein (purified) corresponding to Rat Nestin.
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.|
|Flow Cyt||Use at an assay dependent concentration. PubMed: 21092738
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|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.|
Mesencephalic neural stem cells were washed three times in PBS, fixed in 4% paraformaldehyde for 10 min at room temperature (RT), and permeabilized with PBS plus 0.25% Triton X-100 (PBST) for 10 min at RT (or 15 min at RT if the target antigen was a nucleoprotein). After three washes in PBS, the cells were blocked with 1% BSA for 30 min at RT and incubated with ab6142 staining Nestin (Red) at 1/1000 dilution.
Immunolabeling was visualized by incubation (1 h at RT) with rhodamine-conjugated Affinipure goat anti-mouse or anti-rabbit IgG (1∶100). After three washes in PBS and two in deionized water, immunolabeled cells were counterstained with 50 µl DAPI at 37°C for 10 min and washed in sequence with methanol and ethanol. Cells were observed under a fluorescence microscope.
Fluorescence photomicrographs show that all neurospheres were immunoreactive for the NSCs marker nestin. The neurospheres of peGFPN1-GDNF-transfected mNSCs (termed GDNF-mNSCs) were strong immunopositive for GDNF compare to mNSCs transfected with peGFPN1. Green: FITC, Red: Nestin. Scale bar = 50 µm.
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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