Rat primary antibodies on mouse tissues are
advantageous because rat secondary detection systems can be used with minimum
cross-reactivity to endogenous mouse IgG. Biocare Medical has developed a
Rat-on-Mouse horseradish peroxidase (HRP) polymer detection system with minimal
cross-reactivity to mouse tissues.
It can be used with formalin-fixed paraffin-embedded
tissues, floating sections, frozen sections and cell culture preparations. The
Rat-on-Mouse HRP-Polymer is comprised of two reagents: A Rat Probe that binds to
the rat primary antibody and a Rat-on-Mouse HRP-Polymer that conjugates to the
probe.
This two-step system has been proven to be 10-20 times
more sensitive than conventional conjugated mouse adsorbed anti-rat HRP
secondary detection systems. Biocare Medical has novel products to help
eliminate persistent mouse IgG. XM Factor is a very potent blocker for
eliminating endogenous mouse IgG. By adding 1-2 drops (1 drop equals 33µl) of
the XM Factor to 2.5ml of the Rat-on-Mouse Polymer, endogenous mouse IgG will be
dramatically reduced and/or completely eliminated. Rodent Block M is a blocking
reagent for nonspecific background staining that also reduces endogenous mouse
IgG. Rodent Block M is applied prior to the primary antibody for 15-30 minutes.
Biocare Medical has also developed Rodent Decloaker, an
antigen retrieval solution, which helps reduce and/or eliminate endogenous mouse
and rat IgG and non-specific background staining.
Temperature dependent protocols can be performed using
Biocare’s Decloaking Chamber.