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Native mouse salivary gland NGF purified by modification of the method of Mobley et al (1976). Antigen purity was greater than 95% by PAGE.
Our Abpromise guarantee covers the use of ab6199 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 10 µg/ml.|
|Dot blot||Use at an assay dependent concentration.|
|Neutralising||Use a concentration of 2 - 10 µg/ml. 2ug/ml of ab6199 can neutralize 100ng/ml mouse NGF.|
|WB||Use a concentration of 1 µg/ml.|
|IHC-FoFr||1/300 - 1/5000. (see Abreview on perfusion fixed tissue for detailed protocol).|
|IHC-Fr||Use at an assay dependent concentration.|
ab6199 staining perfusion fixed rat brain and dorsal root ganglion by IHC-Fr. Animals were pre-perfused with Tris buffer pH 10, followed by 4% paraformadehyde and 15% of a saturated solution of picric acid. The brains were post-fixed in the same fixative overnight, cryoprotected in 20% sucrose for 24 hours, frozen and cut with a cryostat. Free floating immunostaining was performed. An Alexa Fluor ® 488 conjugated goat anti-rat antibody was used as the secondary.
The image shows the staining obtained with this antibody using direct fluorescence in the rat cortex and dorsal root ganglion. The staining is not only of the cell body of the cortical neurons but a part of their processes.
ab6199 staining NGF in Human tendon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with Dako FLEX Peroxidase blocking for 5 minutes at room temperature; antigen retrieval was by heat mediation in Dako high pH. Samples were incubated with primary antibody (1/250) for 30 minutes. An undiluted HRP-conjugated Goat polyclonal was used as the secondary antibody.
ab6199 at a 1/500 dilution staining rat brain tissue sections by Immunohistochemistry (Formalin-fixed paraffin-embedded sections). The tissue section was paraformaldehyde fixed and blocked with 2% BSA prior to incubation with the antibody for 24 hours. Bound antibody was detected using a biotinylated goat anti-rabbit IgG antibody.
This image is courtesy of an Abreview submitted by Grazyna Niewiadomska.