Hi, I'm recently working on immunostaining for Pax7 in mouse muscle cryosections. The primary antibody is mouse IgG1 anti-Pax7 (DSHB) and it seems the only antibody working in mouse with a Mouse to Mouse (MOM) IgG blocking reagent. I tried the Vector MOM IgG blocking buffer followed by blocking with 15% goat serum, anti-Pax7 overnight and Invitrogen goat-anti-mouse IgG1 for 30min. It turned out that there was very intense non-specific background. I got this protocol from a Nature paper and also tried different dilution of primary and secondary antibodies but all didn't work. Does anyone have experience with the mouse to mouse kit?
Vector MOM kit is the only way I am aware that has been published to get the DSHB pax7 to work in mouse tissue (to address the previous comment that antibody has been validated by several labs). The DSHB antibody is generally used neat for mouse with the MOM detection kit and can be used diluted 1:3 in human frozen unfixed sections. Our lab had used the 488 supplied with the vector kit as well as Invitrogen 594 for the detection, following the Vector kit protocol. Secondary from invitrogen usually used at 1:500 in PBS buffered with 2% sera. I would stick with the MOM kit. Most researchers that use Pax7 in mouse use that protocol with the vector kit. Also vector is really good at tech support if you email them or call the local rep. Our Rep for McMaster University was really helpful with different Kits and cross-reactions.
I hope that helps
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