Anti-FOX1 | plasma membrane ferroxidase FLP1

Marita Hermann

The FOX1 antibody was tested on whole cell extracts from Chlamydomonas reinhardtii grown both under iron depleted and iron replete conditions. 30 microgram total proteins (quantified by BCA assay) were loaded in each well, separeted on a 13% SDS-PAGE and transferred onto a nitrocellulose membrane. Blocking was performed in PBS 2% milk at room temperature for 1 hour incubation with primary Agrisera FOX1 was also performed in PBS 2% milk by using the suggested 1:1000 dilution. After three washed with PBS 2% milk, secondary anti-rabbit (in PBS 2% milk) was incubated for 1.5 h at a dilution of 1:10,000. Detection was perfomed with a standard ECL assay. Bands of the correct molecular weight could be seen on the film, but background was pretty high and also several aspecific bands were present.

2012-08-16 @ 14:44:42