Can someone recommended antibodies to perform a co-staining (IHC-P) of CD44/CD24 in paraffin embedded sections (mouse samples)?

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Christopher Lange

I assume that you want to stain cancer stem cells (CSCs) with these CD markers, however, you should be aware that even if you add CD326 (ESA, Epcam) to select 2% of (human) the tumor, only 1% of these selected cells are CSCs. See: In measuring the radiosensitivity of breast cancer stem cells (CSCs), cell surface biomarkers purporting to designate a CSC “phenotype” are an insufficient definition of CSCs, since > 98% are not CSCs. Christopher S. Lange, Talal Syed, Paul K. Valestra, Erica Lai, Anna Groysman, Shy’Ann Jie, Manuj Agarwal, Elliot Navo, Robert Rotman, Lisa Dresner, Bozidar Djordjevic. Radiat. Res. 58th Annual Meeting Program Book: pg 126-127, Abstract PS5-48, September 30 – October 3, 2012, San Juan, PR.

For hematological stem cells, the results are probably not much better. You need to use a functional definition as did Till & McCulloch (1961 - 64), but that won't help with fixed paraffin sections. You may need to rethink the plan of your study.

Gudrun Lang

Are these antigenes colocated or on different cells?

If they are on different cells you can make a double-stain with DAB and Red chromogen.

Colocated Antigens are very hard to differentiate with chromogen-IHC. This would be better with immunofluorescence, but autofluorescence of FFPET is high.