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.