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Synthetic peptide based on the amino terminal end of domain-III in the large subunit of Rat Calpain 2. (Peptide available as ab41302.)
Our Abpromise guarantee covers the use of ab39165 in the following tested applications.
|WB||1/1000 - 1/5000. Detects a band of approximately 76 kDa (predicted molecular weight: 80 kDa). Dilution optimised using Chromogenic detection.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 17192410|
|IHC-P||Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
ab39165 (4µg/ml) staining CALPAIN2 in human lung using an automated system (DAKO Autostainer Plus). Using this protocol there is cytoplasmic staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 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.