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Our Abpromise guarantee covers the use of ab26278 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/2000. Fix in 4% paraformaldehyde in 0.1 phosphate buffer, pH 7.4 overnight at 4°C.|
|IHC-Fr||1/2000. Best performance has been seen in frozen sections.|
|Sandwich ELISA||Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Mouse monoclonal [4F3] to GLP1 (ab23472).
We recommend that ab26278 is used as capture antibody. ab26278 binds to the c-terminus and presents the mid-molecular epitope to the ab23472 detection antibody, so it has easy access to it. For the same reason we do not recommend swapping the pair, as the binding of the mid molecular epitope to the coat will not present the c-terminus epitope to the ab26278 antibody in a very favourable manner.
|ELISA||Use at an assay dependent concentration.|
ab26278 (1µg/ml) staining GLP1 in Human pancreas, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.