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mAbs Against HUMAN Protein

4KB51

Antibody Validated
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 labels 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 and in 3/4 cases of acute myeloid leukaemia. While staining was also seen in 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
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
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.