Immunohistochemistry was done on 2-µm paraffin sections. Briefly, the slides were pretreated by boiling for 15 min in a microwave oven in citrate buffer (pH 6). First antibodies specific for MMR proteins and PD-L1 were as follows: MSH2 (clone FE11, dilution 1:100; Calbiochem), MLH1 (clone G168-15, dilution 1:100; BD Pharmingen), MSH6 (1:100; clone EP49; Novus Biologicals), PMS2 (1.50; clone A16-4; BD Pharmingen) and PD-L1 (clone SP263; Ventana Benchmark Ultra, Tuscon). Visualization was carried out following a standard avidin–biotin method (ABC; Vectastain).
Clone ep49
Manufactured by Novus Biologicals
Sourced in United States
The Clone EP49 is a laboratory equipment product. It is used for cell culture and molecular biology applications.
Automatically generated - may contain errors
2 protocols using clone ep49
1
Immunohistochemistry for MMR and PD-L1
2
PD-L1 Expression and Mismatch Repair Assessment
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PD-L1 expression was assessed by IHC staining, using SP263 (Ventana Benchmark Ultra, Tuscon, AZ, USA), the 22C3 pharmDx kit (Agilent Technologies, Santa Clara, CA, USA), or E1L3N XP Rabbit mAb (Cell Signaling Technology, Danvers, MA, USA). If more than 1% of viable tumor cells had PD-L1, they were considered PD-L1-positive [18 (link)]. The tumor proportion score (TPS) was defined as the percentage of viable tumor cells showing partial or complete membrane-staining at any intensity [19 (link)]. The combined positive score (CPS) was defined as the number of PD-L1-positive cells (tumor cells, lymphocytes, and macrophages), divided by the total number of viable tumor cells, and multiplied by 100 [20 (link)].
The microsatellite stability was assessed using antibodies specific for mismatch repair proteins, including mutL homolog 1 (1:10; clone G168-15; BD Pharmingen, San Jose, CA, USA), mutS homolog (MSH)-2 (1:100; clone FE11; Calbiochem, San Diego, CA, USA), MSH6 (1:100; clone EP49; Novus Biologicals, Centennial, CO, USA), and PMS1 homolog 2 (1:50; clone A16-4; BD Pharmingen).
The microsatellite stability was assessed using antibodies specific for mismatch repair proteins, including mutL homolog 1 (1:10; clone G168-15; BD Pharmingen, San Jose, CA, USA), mutS homolog (MSH)-2 (1:100; clone FE11; Calbiochem, San Diego, CA, USA), MSH6 (1:100; clone EP49; Novus Biologicals, Centennial, CO, USA), and PMS1 homolog 2 (1:50; clone A16-4; BD Pharmingen).
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