Anti-Perilipin A antibody (ab3526)

Western blot - Perilipin A antibody (ab3526)

Western blot of perilipin A in 3T3-L1 cell extract using ab3526.

Immunocytochemistry/ Immunofluorescence - Perilipin A antibody (ab3526)

ab3526 staining Perilipin A in rat myofibers by Immunohistochemistry (PFA fixed).

Freshly isolated single myofibers were collected in 0.5 ml DMEM, 5% HS in an Eppendorf tube. Fibers and satellite cells were fixed with 4% paraformaldehyde in phosphate buffered saline, rinsed in PBS and permeabilized with Triton X-100, 0.5% in PBS. After washing, fibers and/or cells were incubated with ab3526 at a 1/50 dilution overnight at 4ºC or 1 hour at 37ºC. Non specific interactions were blocked with 20% goat serum. They were then washed and incubated with labeled secondary antibodies for one hour at room temperature. Satellite cells were then mounted with fluorescent mounting medium plus DAPI 100 ng/ml. Fibers were collected and moved onto a polilysine microscope slide, and then mounted.

Image from Rossi CA et al, PLoS One. 2010 Jan 1;5(1):e8523, Fig 3.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Perilipin A antibody (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.