Mouse Hematopoietic Antigens in Paraffin-embedded Tissues

(Compiled by Dr. JM Ward, NIAID, NIH)

Antigen	Normal Cells Expressing Antigen	Reactive in Formalin-fixed Tissues	Preferred Fixative for Optimal Immunoreactivity	Mouse Tumors Immunoreactive
CD3	Various T lymphocytes	Yes	Bouin's, PF, B5*** Formalin - pressure cooker	T-cell lymphoma
CD45R (B220, Ly-5)	Various B lymphocytes	Yes	Bouin's, PF, B5*** Formalin - microwave, antigen retrieval	B-cell lymphoma
Pax-5	Various B lymphocytes	Yes	MW, EDTA	B-cell lymphoma
Immunoglobulins (IgG, IgA, etc.)	Immunoglobulin-producing B lymphocytes	No	Bouin's, PF, B5*** Formalin - MW H20	B-cell lymphoma
Immunoglobulin human kappa light chains	Immunoglobulin-producing B cells	No	Bouin's, PF, B5*** Formalin -MW H20, overnight incubation, 4C	B-cell lymphoma
Mac-2	Macrophages/ Histiocytes	No	Bouin's***	Histiocytic sarcoma
Lysozyme	Macrophages/ Histiocytes/ Myeloid cells	Yes**	Formalin	Histiocytic sarcoma
Myeloperoxidase	Myeloid cells	Yes	Formalin, Telly's	Myeloid leukemia
NOS2 (nitric oxide synthase)	Myeloid cells	Yes	Formalin, Bouins	Myeloid leukemia
F4/80	Macrophages/ histiocytes	Yes	Formalin - trypsin	Histiocytic sarcoma

Key

PF paraformaldehyde

B cell lymphoma (common lymphoma in B6C3F! mouse also termed lymphoma, lymphoma-mixed or pleomorphic)

*usually requires antigen retrieval or related methods for optimal immunoreactivity. For CD3, some antibodies which are useful in many species may be more optimal in mice

**depends on antibody used

***other quick-fixing preservative can also be used with immediate embedding

Source of Antibodies (our protocols)

CD3 - DAKO, rabbit anti-human, product number A0452, 1:100 on Bouins fixed tissues, 1:400 on formalin fixed tissues with antigen retrieval

CD45R/B220 - Pharmingen, product no. 01122A, biotinylated rat anti-mouse, 1:100 on Bouin's fixed tissues, 1:200 with antigen retrieval.

Pax-5 - Santa Cruz, goat polyclonal C-20, product sc-1974, 1:2000, nuclear antigen

F4/80 - Serotec, rat anti-mouse, product no. mcap497, 1:25, antigen retrieval on formalin fixed tissues.

Human kappa light chains - DAKO rabbit anti-human

Myeloperoxidase - DAKO A0398, formalin - microwave in distilled water, 1:1000

NOS2 (N-20) - Santa Cruz (sc-651) - microwave, citrate buffer, 1:1000

CD45R in Mouse Spleen

Double stain (red = CD3 in PALS, Brown = CD45R in follicle and marginal zone lymphocytes)

Spleen - CD45R in follicles of white pulp

CD45R in follicles of lymph node

CD3 in paracortex of mouse lymph node

CD3 in PALS of mouse spleen

CD3 in mouse Thymic lymphoma

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