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Our Abpromise guarantee covers the use of ab3526 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use at an assay dependent concentration. Can be blocked with Perilipin A peptide (ab5009). By Western blot, this antibody detects an ~62 kDa protein representing Perilipin A from 3T3-L1 cell extract|
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 20049087|
|ICC/IF||Use at an assay dependent concentration. PubMed: 20335253|
Western blot detection of Perilipin A in 3T3-L1 cell extract using ab3526.
ab3526 (4µg/ml) staining perilipin in Human breast adipose using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of adipocytes.
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.
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