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      Biotin Rat Anti-Mouse CD179b
      Biotin Rat Anti-Mouse CD179b
      Two-color analysis of CD179b  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 (Cat. No. 551865, 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.
      Biotin Rat Anti-Mouse CD179b
      Two-color analysis of CD179b  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 (Cat. No. 551865, 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.
      商品详情
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      BD Pharmingen™
      λ5; Lambda 5; VpreB1; Vpreb-2; Immunoglobulin omega chain; Ig omega
      Mouse (QC Testing)
      Rat LEW, also known as Lewis IgG2a, κ
      Purified soluble complex of recombinant µ IgH, λ5, and V[preB]
      Flow cytometry (Routinely Tested)
      0.5 mg/ml
      22363
      AB_394279
      Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
      RUO


      准备和存储

      Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
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      推荐的实验流程

      Because CD179b is expressed at very low levels on bone-marrow-derived early B lineage cells, we recommend the use of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. no. 553141/553142) and amplification of staining by the use of a "bright" second-step reagent, such as Streptavidin-PE (Cat. no. 554061) or Streptavidin-APC (Cat. no. 554067). It is difficult to distinguish the CD179b+ cells among the total bone-marrow population. Therefore, the staining protocol described by Winkler, et al., should be used. Essential features of this procedure include immunomagnetic depletion of surface IgM+ cells and incubation of the cells at 37°C for 1 hour in tissue culture medium.

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      商品通知

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      551865 Rev. 4
      抗体详情
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      LM34

      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.

      551865 Rev. 4
      格式详情
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      Biotin
      Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
      Biotin
      551865 Rev.4
      报价单和参考
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      研发参考 (9)

      1. Karasuyama H, Rolink A, Melchers F. A complex of glycoproteins is associated with VpreB/lambda 5 surrogate light chain on the surface of mu heavy chain-negative early precursor B cell lines. J Exp Med. 1993; 178(2):469-478. (Immunogen). 查看参考
      2. Karasuyama H, Rolink A, Shinkai Y, Young F, Alt FW, Melchers F. The expression of Vpre-B/lambda 5 surrogate light chain in early bone marrow precursor B cells of normal and B cell-deficient mutant mice. Cell. 1994 April; 77(1):133-143. (Biology). 查看参考
      3. Martensson IL, Ceredig R. Review article: role of the surrogate light chain and the pre-B-cell receptor in mouse B-cell development. Immunology. 2000; 101(4):435-441. (Biology). 查看参考
      4. Meffre E, Papavasiliou F, Cohen P, et al. Antigen receptor engagement turns off the V(D)J recombination machinery in human tonsil B cells. J Exp Med. 1998; 188(4):765-772. (Biology). 查看参考
      5. Melchers F, ten Boekel E, Seidl T, et al. Repertoire selection by pre-B-cell receptors and B-cell receptors, and genetic control of B-cell development from immature to mature B cells. Immunol Rev. 2000; 175:33-46. (Biology). 查看参考
      6. Ohnishi K, Shimizu T, Karasuyama H, Melchers F. The identification of a nonclassical cadherin expressed during B cell development and its interaction with surrogate light chain. J Biol Chem. 2000; 275(40):31134-31144. (Biology). 查看参考
      7. Rolink A, Grawunder U, Winkler TH, Karasuyama H, Melchers F. IL-2 receptor alpha chain (CD25, TAC) expression defines a crucial stage in pre-B cell development. Int Immunol. 1994; 6(8):1257-1264. (Biology). 查看参考
      8. Shimizu T, Mundt C, Licence S, Melchers F, Martensson IL. VpreB1/VpreB2/lambda 5 triple-deficient mice show impaired B cell development but functional allelic exclusion of the IgH locus. J Immunol. 2002; 168(12):6286-6293. (Biology). 查看参考
      9. Winkler TH, Rolink A, Melchers F, Karasuyama H. Precursor B cells of mouse bone marrow express two different complexes with the surrogate light chain on the surface. Eur J Immunol. 1995; 25(2):446-450. (Biology). 查看参考
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      551865 Rev. 4

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      Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

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