mAbs Against HUMAN Protein
4KB51
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | LRF Haemato-oncology Group, University of Oxford |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | 4KB51 |
| PROTEIN: | - |
| ANTIGEN USED: | Hairy cell leukaemia |
| FUSION PARTNER: | NS1 |
| ISOTYPE: | IgG2a |
| SPECIES REACTIVITY: | human |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
| APP RECOMMENDED: | IHQ-frozen, IP, Flow cytometry, WB |
| APP NO RECOMMENDED: | IHQ-paraffin |
| APP NO TESTED: | IF |
Description
The 4KB51 antigen is an intracellular marker. The antibody labelled B- and T-cells but not germinal centre cells or plasma cells. Labelling was also observed of suprabasal epithelial cells. Staining was observed in 11 out of 11 cases of hairy cell leukaemia studied and in the neoplastic cells in 2 cases of chronic myeloid leukaemia, 3/4 cases of acute myeloid leukaemia and 4/16 cases of mantle cell lymphoma. All other B-cell tumours (CLL, CALL, Myeloma, DLBCL), T-cell tumours (ALCL, ALL, PTCL) and HL studied were unlabelled.
References
Bryan N & Pulford K. 2002 Heterogeneity of expression of the 4KB51 antigen on cord blood and peripheral blood cells. In Proceedings of VIIth International Workshop on White Cell Differentiation Antigens. Ed Mason DY et al. 2002. Oxford University Press, Oxford. P.137-139.
Pulford K, Micklem KJ, Jones M, Pezzella F, Mayne KM, Morrison H, Thomas J, Falini B, Norton A, Mason DY. A novel internal antigen which distinguishes germinal centre cells from other B-cell types. Immunology 1994; 82:154-63.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 4KB51 | Neat | Supernatant | None | HRP or APAAP | Tonsil / No first ab | intracellular | Unknown | |
| Paraffin tissue | |||||||||
Not recommended Reagent discarded for this application | Not reactive | / | |||||||
| Immunofluorescence | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 4KB51 | Neat | / | ||||||
Enlarge image
- Immunoperoxidase labelling of tonsil and thymus to show the distribution of the 4KB51 antigen
- Antibody 4KB51 labels the mantle zone and interfollicular areas of tonsil while germinal centre cells are unlabelled. Labelling in the thymus is largely restricted to a subpopulation of cells in the medulla.
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
| Immunoprecipitation | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | 4KB51 | Neat | supernatant | / mouse IgG2a | ||||
Enlarge image
- Biochemical characterisation of the 4KB51 antigen.
- Immunoaffinity purified 4KB51 was biotinylated an immunoprecipitated with antibody coated Prot G-Sepharose before SDS-PAGE. After Western blotting with Steptavidin-HRP, a) the 4KB51 antigen was 160 kD (arrowhead, unreduced) and c) 68 kD (reduced). Lanes b) and c) show the results obtained using isotype matched controls.