Table 2.

Antibodies used for immunohistochemistry and EIA; GdC15, GdQ13 and GdN20 were purchased from SantaCruz Biotechnologies, Santa Cruz, CA (SCBT)

Immunohistochemistry
Antibody	Epitope	Source/ Clonality	Epitope retrieval	Blocking	Dilution	Incubation	Reaction system used for detection
GdC15 (SCBT)	C-terminal	goat/polyclonal	Proteinase K (30 min, RT)	UV-Block (45 min)	1:1000 (UV Block)	30 min (RT)	Vectastain elite kit (goat IgG)
GdQ13 (SCBT)	C-terminal	goat/polyclonal	citrate buffer (pH 6, 5 min, pressure cooker)	UV-Block (45 min)	1: 300 (UV Block)	o.n. (4°C)	Vectastain elite kit (goat IgG)
GdA (Jeschke et al. 2006)	Mixed glycan/protein epitope	mouse/ monoclonal	citrate buffer (pH 6, 5 min, pressure cooker)	1.5% horse serum (20 min)	1:3000 (DAKO dil.)	o.n. (4°C)	Vectastain elite kit (mouse IgG)
Competitive enzyme immuno assay (EIA)
Antibody	Epitope	Source/ Clonality	Competing peptide	Blocking	Dilution	Incubation	Reaction system used for detection
GdQ13 (SCBT)	C-terminal	goat/ polyclonal	GdQ13-P (SCBT)	1% BSA, 0.5% NaN3 in PBS	1.5 ng/ml	1 hour (RT)	POD-strepavidin
GdN20 (SCBT)	N-terminal	goat/ polyclonal	GdN20-P (SCBT)	1% BSA, 0.5% NaN3 in PBS	6 ng/ml	1 hour (RT)	POD-strepavidin

rt = room temperature, BSA = bovine serum albumin, PBS = phosphate buffered salt solution, o.n. = over night, POD = horse radish peroxidase.

Competing peptides came from SCBT as well and were biotinylated by the manufacturer; unmodiefied GdQ13-P, GdN20-P were used for setting up the standard curve. Preparation of GdA is published in Jeschke et al. 2006. Epitope retrieval: Proteinase K was from Quiagen (Hilden, Germany), citrate buffer contained 0.1 M citric acid and 0.1 M sodium citrate in distilled water (pH 6.0). Blocking: UV-Block was purchased from Thermo Scientific (Bonn, Germany) and horse serum was a component of the Vectastain elite kit (Vector Laboratories, Burlingame, USA).