Table 3 Antibodies used for... | American Society of Hematology

Table 3

Antibodies used for immunohistochemistry in paraffin sections

Antibody	Targets	Dilution	Antigen retrieval treatment	Detection System from DAKO
Anti–PD-1	T_FH cells	1:5	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer, pH 6	Strept. AP system
Anti-CD3	T cells	1:100	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer, pH 6	APAAP system
Anti-CD21	FDCs and MZ analog B cells	1:50	20 minutes with Pronase at 37°C	APAAP system
Anti-BCl6	T_FH cells and GC B cells	1:50	Heated in high-pressure cooker for 5 minutes in sodium citrate buffer, pH 6	Strept. AP system
Anti-Ki67	Proliferating cells	1:2000	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer, pH 6	APAAP system
Anti-CD23	FDCs and MZ analog B cells	1:20	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer, pH 6	APAAP system
Anti-IgM	Plasma cells and B cells i	1:1000	Heated in high-pressure cooker for 5 minutes in sodium citrate buffer, pH 6	Strept. AP system
Anti-GPIV	Platelet glycoprotein IV: a target of autoantibody in ITP patients.	1:200	Heated in high-pressure cooker for 1 minute in EDTA	Strept. AP system
Anti-Foxp3	T(reg)	1:50	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer pH6 and 10 minutes 3% H₂O₂ blocking for endogenic peroxidase	EnVision+ System

Antibody	Targets	Dilution	Antigen retrieval treatment	Detection System from DAKO
Anti–PD-1	T_FH cells	1:5	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer, pH 6	Strept. AP system
Anti-CD3	T cells	1:100	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer, pH 6	APAAP system
Anti-CD21	FDCs and MZ analog B cells	1:50	20 minutes with Pronase at 37°C	APAAP system
Anti-BCl6	T_FH cells and GC B cells	1:50	Heated in high-pressure cooker for 5 minutes in sodium citrate buffer, pH 6	Strept. AP system
Anti-Ki67	Proliferating cells	1:2000	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer, pH 6	APAAP system
Anti-CD23	FDCs and MZ analog B cells	1:20	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer, pH 6	APAAP system
Anti-IgM	Plasma cells and B cells i	1:1000	Heated in high-pressure cooker for 5 minutes in sodium citrate buffer, pH 6	Strept. AP system
Anti-GPIV	Platelet glycoprotein IV: a target of autoantibody in ITP patients.	1:200	Heated in high-pressure cooker for 1 minute in EDTA	Strept. AP system
Anti-Foxp3	T(reg)	1:50	Heated in high-pressure cooker for 2 minutes in sodium citrate buffer pH6 and 10 minutes 3% H₂O₂ blocking for endogenic peroxidase	EnVision+ System

T_FH indicates T follicular helper; FDC, follicular dendritic cell; MZ, marginal zone; GC, germinal center; and ITP, immune thrombocytopenia.