The pre-B cell receptor (pre-BCR) expressed during the early stages of B lymphocyte development is a heterodimer of immunoglobulin heavy chain (IgH) with surrogate light chain, which is an Ig-light-chain-like molecule composed of the non-covalently linked CD179b (λ5) and CD179a (VpreB) proteins. The pre-BCR is believed to control IgH repertoire selection and proliferation of differentiating B lymphocytes. The LM34 antibody reacts with λ5 and surrogate light chain, but not VpreB alone, in transfected X63-Ag8.653 cells. It detects surrogate light chain on pro-B and pre-B cell lines, in the presence or absence of IgH, but not on IgM-positive B lymphocytes. It also detects surrogate light chain or λ5 on the surface and in the cytoplasm of pro-B or pre-B cells from the bone marrow of normal or RAG2-deficient mice. It has been noted that the (CD45R/B220) cell-surface expression of surrogate light chain is upregulated after a one-hour incubation of bone-marrow leukocytes in tissue culture medium at 37 °C. At the earliest stages of B lymphopoiesis, before IgH is available, the surrogate light chain associates with a complex of glycoproteins, including a nonclassical cadherin, which could be involved in selective adhesion events during B-lymphocyte development. After immunization, cell-surface λ5 is detected on a subset of splenic Ig light chain-positive germinal-center B lymphocytes. Two-color analysis of CD179b (?5) expression on bone-marrow pre-B lymphocytes. After immunomagnetic depletion of IgM+ cells using biotinylated anti-mouse IgMb mAb AF6-78 (Cat. no. 553519), C57BL/6 bone-marrow leukocytes were incubated for 1 hour in DMEM at 37 °C to enhance surrogate light chain expression. Then the leukocytes were stained with either biotinylated rat IgG2a, κ isotype control mAb R35-95 (Cat. no. 553928, left panel) or biotinylated mAb LM34 (right panel) in the presence of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. no. 553141/553142), followed by Streptavidin-APC (Cat. no. 554067) and FITC-conjugated anti-mouse CD45R/B220 mAb 9 RA3-6B2 (Cat. no. 553087/553088). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
BD Pharmingen™ Biotin Rat Anti-Mouse CD179b - Biotin - Clone LM34 - Isotype Rat IgG2a, κ - Reactivity Ms - 0.1 mg
Purification
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Immunogen
Purified soluble complex of recombinant mu IgH, lambda5, and V[preB]
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
Storage
4 °C
Storage Comment
Store undiluted at 4°C.
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