Is an antibody not specific or Western blot protocol not optimized?

Courtesy of Dr. Lea Vojta and Dr. Hrvoje Fulgosi, Laboratory for Molecular Plant Biology and Biotechnology, Division of Molecular Biology, Institute Ruđer Bošković, Croatia.

Antibody: TROL | Thylakoid rhodanese-like protein, AS19 4257.

Western blot protocol before optimization
Protein load/well: 20 µg
Blocking: 5 % milk
Primary antibody: 1: 2000 ON/4ºC
Secondary antibody: 1: 50 000 1h/RT

Western blot protocol after optimization
Protein load/well: 2 µg
Blocking: 5 % milk
Primary antibody: 1: 2000 ON/4ºC
Secondary antibody: 1: 50 000 1h/RT

Visible improvement was achieved with decreased protein load/well (10x).

In a need to optimize your western blot protocol?
Request a copy of the Agrisera Western blot trouble shooting brochure, or send your inquiry directly to [email protected].


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