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Biotin Rat Anti-Mouse Pre-B Cell Receptor
Biotin Rat Anti-Mouse Pre-B Cell Receptor

Two-color analysis of Pre-B cell receptor (Pre-BCR) expression on bone-marrow pre-B lymphocytes. After immunomagnetic depletion of IgM+ cells using biotinylated anti-mouse IgM[b] mAb AF6-78 (Cat. No. 553519), C57BL/6 bone-marrow leukocytes were incubated for 1 hour in DMEM at 37°C to enhance Pre-BCR 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 SL156 (Cat. No. 551863, 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 (Cat. No. 553087/553088). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Two-color analysis of Pre-B cell receptor (Pre-BCR) expression on bone-marrow pre-B lymphocytes. After immunomagnetic depletion of IgM+ cells using biotinylated anti-mouse IgM[b] mAb AF6-78 (Cat. No. 553519), C57BL/6 bone-marrow leukocytes were incubated for 1 hour in DMEM at 37°C to enhance Pre-BCR 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 SL156 (Cat. No. 551863, 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 (Cat. No. 553087/553088). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Pharmingen™
Pre-BCR
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2a, κ
Purified soluble complex of recombinant µ IgH, λ5, and VpreB
Flow cytometry (Routinely Tested)
0.5 mg/ml
AB_394277
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

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.

Recommended Assay Procedures

Because the pre-BCR 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 dinstinguish the cells expressing pre-BCR among the total bone-marrow population. Therefore, the staining protocol is described in the literature. 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.

Product Notices

  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.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
551863 Rev. 3
Antibody Details
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SL156

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 Lambda 5 (λ5) and VpreB proteins. The pre-BCR is believed to control IgH repertoire selection and proliferation of differentiating B lymphocytes. The SL156 antibody reacts with a conformational epitope of the pre-BCR in transfected X63-Ag8.653 cells but not with surrogate light chain, λ5, or VpreB in the absence of IgH. It detects pre-BCR on pre-B cell lines, but not on pro-B cell lines or IgM-positive splenocytes. It also detects pre-BCR on the surface of pre-B cells, but not on Ig light chain (IgL)-positive B lymphocytes, from the bone marrow of normal mice. It has been noted that the cell-surface expression of pre-BCR is upregulated after a one-hour incubation of bone-marrow leukocytes in tissue culture medium at 37°C. After immunization, cell-surface pre-BCR is detected on a subset of splenic IgL+ germinal-center B lymphocytes.

551863 Rev. 3
Format Details
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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
551863 Rev.3
Citations & References
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Development References (6)

  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). View Reference
  2. 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). View Reference
  3. 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). View Reference
  4. 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). View Reference
  5. 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). View Reference
  6. 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. (Immunogen). View Reference
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551863 Rev. 3

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