Comparative Medicine Branch Veterinary Pathology
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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