While benign (reactive) B-lymphocytic populations produce Ig-molecules containing an almost equal amount of kappa and lambda light chains, i.e. the number of cells producing kappa is more or less equal to the number producing lambda, neoplastic B-lymphocytic populations has light chain restriction (i.e., are monoclonal) producing either kappa or lambda. Consequently, demonstration of light chains is the most important procedure in the diagnosis of neoplasms of B-lymphocytes (lymphomas and leukemias). The amount of Ig produced in individual types of B-lymphocytic neoplasia vary and is often small, demanding a sensitive technique for detection. About 80% of non-Hodgkin lymphomas are B-cell lymphomas, of which the large majority express surface (s) IgM, although the expression in small cell lymphoma/chronic lymphocytic leukaemia is weak. Plasmacytoma/multiple myeloma do not show sIgM. Lymphoplasmacytic lymphoma most often shows sIgM, while plasmacytoma/multiple myeloma show sIgG in 50% and sIgA in 20%. Light chain restriction is the single most important marker for neoplasms of B-lymphocytic origin. Demonstration of heavy chains can sometime be of aid in the study of malignant lymphomas, as lymphoplasmacytic lymphomas are usually focally IgM positive, while plasmacytoma/multiple myeloma in most cases express strong cytoplasmic IgG or IgA. Precursor B-cell neoplasms are are Ig negative. Normal tonsil is appropriate control tissue: approximately 50% of the mantle zone B-cells should show a distinct membrane staining reaction, while the rest should be unstained.
Background References
1. Kato A. et. al. An expanded genetic code facilitates antibody chemical conjugation involving the lambda light chain. Biochem Biophys Res Commun. 2021 Mar
2. van der Kant R. et. al. Adaption of human antibody λ and κ light chain architectures to CDR repertoires. Protein Eng Des Sel. 2019 Dec
Western blot analysis of Lambda Light Chain on human small intestine tissue lysates with Rabbit anti-Lambda Light Chain antibody (HA721414) at 1/1,000 dilution.
Lysates/proteins at 20 µg/Lane.
Predicted band size: 11 kDa Observed band size: 25 kDa
Exposure time: 2 minutes;
12% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721414) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Lambda Light Chain antibody (HA721414) at 1/3,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721414) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Application: IF-Tissue
Species: Human
Site: tonsil
Sample: Paraffin-embedded section
Antibody concentration: 1/1,000
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