Hello All,

I am staining for human extracellular matrix proteins using mouse
anti-collagen/fibronectin/elastin etc. I am using a Fluorscein-conjugated
goat anti-mouse secondary antibody. Despite various preventative methods I
am getting perfect staining on my negative (or null) control which has no
primary added.

I've used various dilutions, Bovine, human and rabbit serum block, and
multiple blocking steps

all with no improvement. If anything, the secondary has greater affinity
for matrix proteins than it does for the IgG part of the primary. I'm
wondering whether, since the serum itself has matrix proteins in it,
whether to completely forego the blocking step.


Any thoughts or ideas extremely welcome,

Thanks

Gail



Gail Donegan
Department of Clinical Engineering,
The University of Liverpool,
Duncan Building,
Daulby Street,
Liverpool,
L69 3GA.
U.K.

Tel: +44(0)151 706 5198
Fax: +44(0)151 706 5803

.

Gail Donegan
Department of Clinical Engineering,
The University of Liverpool,
Duncan Building,
Daulby Street,
Liverpool,
L69 3GA.
U.K.

Tel:    +44(0)151 706 5198
Fax:    +44(0)151 706 5803