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Our Abpromise guarantee covers the use of ab8448 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/100 - 1/300.|
|WB||1/1000. at this dilution, the antibody will strongly detect approximately 250 ng of OPN protein on a blot.|
|IHC-Fr||1/100 - 1/500. PubMed: 16128620|
ab8448 staining Osteopontin in mouse developing skeleton tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with paraformaldehyde before permeabilization with 0.1% Triton and blocking with 20% serum was performed for 1 hour at RT. The sample was incubated with primary antibody (1/200) in 20%FBS/PBS for 16 hours at 40C. An Alexa Fluor®488-conjugated donkey polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution. In the image: Red Rhodamine Phalloidin (muscle), Blue DAPI (nuclei), Green Osteopontin.
Breast tumour section. Osteopontin is a normal component of elastic fibers in the breast (shown heavily stained in this section). There is also weak staining of the extracellular matrix. Osteopontin is not believed to be expressed inside breast tumour cells, and there is no staining in the intracellular region of the breast cells in this section.
Osteopontin antibody (ab8448) used at 1:100-1:300. No antigen retrieval is required.
ab8448 staining Osteopontin in Mouse femur tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% BSA for 1 hour at 35°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/50) for 14 hours at 4°C. An Alkaline Phosphatase-conjugated Goat anti-rabbit IgG F(ab')2 polyclonal (1/100) was used as the secondary antibody.