Immunohistowax processing, a new fixation and embedding method for light microscopy, which preserves antigen immunoreactivity and morphological structures: visualisation of dendritic cells in peripheral organs

Table 2

Antigens successfully detected on Immunohistowax processed mouse tissue sections

Antigens	Clones	Specificity	Dilution	Sources
*Fusion protein; °synthesised hapten carrier protein.
Ars, arsenate; BSA, bovine serum albumin; DC, dendritic cell; h, human; IFN-γ, interferon γ; Ig, immunoglobulin; IL, interleukin; L, ligand; MHC, major histocompatibility complex; MΦ, macrophage; NK, natural killer; PMN, polymorphonuclear cell; TCR, T cell receptor; VCAM, vascular cell adhesion molecule.
CD2	12–15	Cell adhesion molecule	10–20 μg/ml	Southern Biotechnology Associates, Inc
CD3	7D6	T cells	20–30 μg/ml	Coulie and colleagues¹⁴
CD4	RM4–5	T helper cells	20 μg/ml	Pharmingen
CD8	53–6.7	T cells	20 μg/ml	Pharmingen
CD90/Thy 1.2	53–2.1	T cells	10–20 μg/ml	Pharmingen
TCRαβ	H57–597	T cells (TCR)	20–30 μg/ml	Pharmingen
TCRγδ	13D5	T cells (TCR)	20–30 μg/ml	Peterman and colleagues¹⁵
TCR Vβ8	F23.2	T cells (TCR)	20 μg/ml	ATCC
TCR Vβ6	44.22.1	T cells (TCR)	20 μg/ml	Payne and colleagues¹⁶
B220/CD45R	RA3–6B2	B cells	5–10 μg/ml	Pharmingen
IgD	LoMD6	B cells	5–10 μg/ml	Lefebvre and colleagues¹⁷
IgM	LoMM9	B cells	5–10 μg/ml	Lefebvre and colleagues¹⁷
MHC-II	14.4.4	B cells, DC, Mφ	5–10 μg/ml	ATCC
CD11c	N418	Mainly DC	5–10 μg/ml	Metlay and colleagues¹⁸
CD11c	HL3	Mainly DC	5–10 μg/ml	Pharmingen
CD11b/Mac-1	M1/70	MΦ	5–10 μg/ml	Pharmingen
NK	5E6	NK cells	10–20 μg/ml	Pharmingen
Pan-NK	DX5	NK cells	10–20 μg/ml	Pharmingen
Ly-6G/GR1	RB6–8C5	Granulocytes	10–20 μg/ml	Pharmingen
PMN antigen	7/4	Neutrophils	10–20 μg/ml	Caltag
CD40	3/23	B cells, DC, Mφ	10–20 μg/ml	Hasbold and colleagues¹⁹
Intracytoplasmic granules	2A1	Mature DC	5–10 μg/ml	Inaba and colleagues²⁰
Intracytoplasmic granules	M342	Mature DC	Undiluted supernatant	Agger and colleagues²¹
DEC-205	NLDC-145	Mature DC	20–30 μg/ml	Kraal and colleagues²²
CD35	8C12	CR1 (Follicular DC)	5–10 μg/ml	Pharmingen
CD86/B7.2	GL1	Activated B cells, DC, Mφ	10–20 μg/ml	Hathcock and colleagues²³
CD25	7D4	Activated T cells, B cells, DC	10–20 μg/ml	Pharmingen
Ly-77	GL7	Activated T cells, B cells	10–20 μg/ml	Pharmingen
CD44	1M7	Activated T cells	10–20 μg/ml	Pharmingen
CD69	H1.2F3	Activated T cells	10–20 μg/ml	Pharmingen
Phosphotyrosin	4G10	Activated cells	20–30μg/ml	Upstate Biotechnology Incorporation
OX40	mOX40L–hIgG1*	Activated T cells	20–30μg/ml	Cantab Pharmaceutical Research Ltd
VCAM-1	M/K-2	Adhesion molecule	10–20 μg/ml	R&D Systems
Anti-Ars Ig	Ars-BSA°	Ars specific B cells	20 μg/ml
IL-2	S4B6	Cytokine	20–30 μg/ml	Pharmingen
IL-4	11B11	Cytokine	20–30 μg/ml	Pharmingen
INF-γ	Db1	Cytokine	20–30 μg/ml	Paineau and colleagues²⁴
IL-10	JESS-2A5	Cytokine	20–30 μg/ml	Pharmingen

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Immunohistowax processing, a new fixation and embedding method for light microscopy, which preserves antigen immunoreactivity and morphological structures: visualisation of dendritic cells in peripheral organs

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