For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome.
Chicken gizzard actin.
Our Abpromise guarantee covers the use of ab3280 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1-2µg for 106 cells.|
|IP||Use at 2 µg/mg of lysate. Use Protein G. This has been tested in IP on "denatured" cell lysate (treated with 5% SDS).|
|WB||Use a concentration of 0.5 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).|
|IHC-P||Use a concentration of 0.5 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use at an assay dependent dilution. See Bryce et al. Fix cells with 4% paraformaldehyde and permeabilize with chilled methanol.|
Lanes: 1-3: 4oC (x1 freeze/thaw)
Lanes: 4-6: 4oC (x2 freeze/thaws)
Lanes 7-9: 4oC (x4 freeze/thaws)
This image is courtesy of an anonymous AbreviewBlocking Step: 5% Milk for 30 minutes at 25°C.
ab3280 (1µg/ml) staining actin in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining throughout the tissue.
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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"