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Our Abpromise guarantee covers the use of ab6214 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-FoFr||1/1000. PubMed: 16052497
Following perfusion fixation with 4% PFA, cryoprotect the tissue in 20% sucrose overnight. Cut on cryostat sections for free floating IHC. Incubate primary antibody for 48 hrs at 4°C.
|IHC-Fr||Use at an assay dependent concentration.|
ab6214 staining Pan Orexin in mouse brain tissue sections by Immunohistochemistry (PFA fixed free floating sections). Tissue samples were fixed with paraformaldehyde, permeablized with 1% Triton X-100 and blocked with 1% BSA for 10 minutes at room temperature. The sample was incubated with primary antibody (1/2000 in 1X PBS + 0.1% Triton X-100 + 1% BSA) at 4°C for 24 hours. An Alexa Fluor® 594-conjugated donkey anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.
Immunohistochemical staining of orexin A in rat lateral hypothalamus useing Anti-Orexin A antibody (ab6214) at 1:50000 dilution. The primary antibody was detected with the biotinylated secondary antibody-ABC method and nickel-diaminobenzidine chromogen.
Immunohistochemical staining of orexin A in rat zona incerta neurons using Rabbit antibody to Orexin A (ab6214) at a dilution of 1 in 1000.