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)
![CD45R in Mouse Spleen](https://webarchive.library.unt.edu/eot2008/20081014101943im_/http://www3.niaid.nih.gov/NR/rdonlyres/C8D1067F-AFBB-46A0-8022-145A56E57E04/0/cd5cd45r.jpg)
Spleen - CD45R in follicles of white pulp
![Spleen - CD45R in follicles of white pulp](https://webarchive.library.unt.edu/eot2008/20081014101943im_/http://www3.niaid.nih.gov/NR/rdonlyres/AAC1EA96-ACDF-4D0C-9028-2274DB0504B6/0/b220low.jpg)
CD45R in follicles of lymph node
![CD45R in follicles of lymph node](https://webarchive.library.unt.edu/eot2008/20081014101943im_/http://www3.niaid.nih.gov/NR/rdonlyres/9D6744B4-D23D-44EE-BFB4-6DEFE8259124/0/b220ln.jpg)
CD3 in paracortex of mouse lymph node
![CD3 in paracortex of mouse lymph node](https://webarchive.library.unt.edu/eot2008/20081014101943im_/http://www3.niaid.nih.gov/NR/rdonlyres/66AE2F57-9861-42E6-93E9-9C4A5F574581/0/cd3ln.jpg)
CD3 in PALS of mouse spleen
![CD3 in PALS of mouse spleen](https://webarchive.library.unt.edu/eot2008/20081014101943im_/http://www3.niaid.nih.gov/NR/rdonlyres/07F272C2-528B-4D91-B1CB-9CF4643C78DB/0/cd3sp.jpg)
CD3 in mouse Thymic lymphoma
![CD3 in mouse Thymic lymphoma](https://webarchive.library.unt.edu/eot2008/20081014101943im_/http://www3.niaid.nih.gov/NR/rdonlyres/3C4DDE9A-EA5D-4709-AB1A-C7D806140A4F/0/cd3thymicl.jpg)
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