Table 2.

Overview of antibody and chemicals used in the staining process

Sialyl Lewis X	Sialyl Lewis A	Lewis Y
Blocking solutiona: 20 min	Blocking solutionb: 5 min	Blocking solutionc: 3 min
Primary antibodyd	Primary antibodyd	Primary antibodyd
1:200 in PBSe	1:80 in PBSe	1:50 in PBSe
Incubation 16 h, 4 °C	Incubation 60 min, 23 °C	Incubation 16 h, 4 °C
Secondary antibodyf	PostBlocak: 20 min	Secondary antibodyf
1:50 in DAKO diluentg		1:200 in DAKO diluentg
Incubation 30 min, 23 °C		Incubation 30 min
ABC-complexh: 30 min	HRP Polymera: 30 min	ABC-complexh: 30 min
Chromogen: DABi, 47 s	Chromogen: DABi, 43 s	Chromogen: DABi, 4 min

^aVectastain Elite ABC Kit, diluted NORMAL serum

^bZytoChem Plus HRP Polymer Kit (Mouse/Rabbit) 3 × 100, Cat.No. POLHRP-100

^cUniversal Blocking Reagent (10X), REF HK085-5KE

^dInformation in Table 3

^eDulbecco’s phosphate-buffered saline

^fBiotinylated Goat-anti-Mouse IgM, Linaris, Nr. ZMB2020

^gDAKO Antibody Diluent with Background Reducing components, REF S3022

^hVectastain Elite ABC Kit, REAGENT A (Avidin, ABC Elite) 30,005, REAGENT B (Biotinylated HRP, ABC Elite) 30,006

ⁱLiquid DAB + Substrate Chromogen System, REF K3468