We tested by western blot a rabbit polyclonal PPARγ Antibody (H-100) cat# sc-7196. It is raised against an epitope corresponding to amino acids 8-106 mapping at the N-terminus of PPARγ1 of human origin. As shown in figure, this antiserum detects (@1:100; 40 microg proteins) multiple bands in brown adipose tissue and brain lysates from the mouse, but no clear band at the expected molecular size.
Using a standard protocol, the proteins were extracted by homogenizing fat and brain samples in lysis buffer, resolved by SDS-PAGE and then transferred to a nitrocellulose membrane by electroblotting (100 V for 1 to 2 hours). The membrane was incubated with the primary antiserum overnight at 4°C and with the secondary antibody for 1 hour at room temperature.
On May 28, 2016 Laurent Gautron, UT Southwestern wrote:
We tested by western blot a rabbit polyclonal PPARγ Antibody (H-100) cat# sc-7196. It is raised against an epitope corresponding to amino acids 8-106 mapping at the N-terminus of PPARγ1 of human origin. As shown in figure, this antiserum detects (@1:100; 40 microg proteins) multiple bands in brown adipose tissue and brain lysates from the mouse, but no clear band at the expected molecular size.
Using a standard protocol, the proteins were extracted by homogenizing fat and brain samples in lysis buffer, resolved by SDS-PAGE and then transferred to a nitrocellulose membrane by electroblotting (100 V for 1 to 2 hours). The membrane was incubated with the primary antiserum overnight at 4°C and with the secondary antibody for 1 hour at room temperature.