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FITC Rat Anti-Mouse CD93 (Early B Lineage)
FITC Rat Anti-Mouse CD93 (Early B Lineage)

Three-color analysis of the expression of the CD93 (Early B Lineage) antigen in mouse bone marrow. BALB/c bone marrow leukocytes were simultaneously stained with PE Rat anti-Mouse CD45R/B220(Cat. No. 553089/553090), APC Rat Anti-Mouse IgM (Cat. No. 550676), and either FITC Rat IgG2b, κ isotype control (Cat. No. 553988) or FITC Rat anti-Mouse CD93 (Early B Lineage) (Cat. No. 559156/561990). Live cells were selected by exclusion of propidium iodide. Double-positive IgM+ CD45R+ cells were gated (left panel) and analyzed for FITC signal. In the right panel, the solid histogram represents the isotype control, while the open histogram represents the expression of the Early B Lineage antigen. Approximately 70% of the IgM+ CD45R+ bone marrow cells are also AA4.1 positive. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Three-color analysis of the expression of the CD93 (Early B Lineage) antigen in mouse bone marrow. BALB/c bone marrow leukocytes were simultaneously stained with PE Rat anti-Mouse CD45R/B220(Cat. No. 553089/553090), APC Rat Anti-Mouse IgM (Cat. No. 550676), and either FITC Rat IgG2b, κ isotype control (Cat. No. 553988) or FITC Rat anti-Mouse CD93 (Early B Lineage) (Cat. No. 559156/561990). Live cells were selected by exclusion of propidium iodide. Double-positive IgM+ CD45R+ cells were gated (left panel) and analyzed for FITC signal. In the right panel, the solid histogram represents the isotype control, while the open histogram represents the expression of the Early B Lineage antigen. Approximately 70% of the IgM+ CD45R+ bone marrow cells are also AA4.1 positive. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Pharmingen™
Aa4; Cd93; C1qRp; C1qr1; Ly-68; Ly68; C1q/MBL/SPA receptor
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
Pre-B lymphoma 70Z/3, derived from (C57BL/6 x DBA/2)F1 mouse
Flow cytometry (Routinely Tested)
0.5 mg/ml
AB_397197
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with FITC under optimum conditions, and unreacted FITC was removed.

Recommended Assay Procedures

For detection of the CD93 (Early B Lineage) antigen in the spleen, we recommend amplification of the staining signal through the use of Biotin Rat Anti-Mouse CD93 (Early B Lineage) (Cat. No. 550434), followed by a "bright" second-step reagent, such as Streptavidin-PE (Cat. No. 554061).

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.
559156 Rev. 8
Antibody Details
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AA4.1

The AA4.1 monoclonal antibody specifically recognizes the Early B Lineage antigen which is also known as CD93, AA4 antigen, Ly-68, and Complement component C1q receptor (C1qRp). This 130-140-kDa type I transmembrane glycoprotein is expressed on immature B lymphocytes in the adult bone marrow and on  hematopoietic progenitors and stem cells in adult bone marrow, fetal liver, and embryonic yolk sac. Although CD93+ cells are most plentiful in adult mouse bone marrow, a smaller number of CD93+ cells which express lower CD93 levels can be detected in the adult spleen using bright fluorescent conjugates of the AA4.1 antibody or an amplified indirect immunofluorescent staining procedure. It has been observed that the staining pattern of the 493 monoclonal antibody is similar to that of the AA4.1 antibody, in that both antibodies precipitate molecules of the same molecular weight. Staining with the AA4.1 antibody is not blocked by the 493 antibody. These results suggest that the antibodies recognize separate epitopes on the same Early B Lineage antigen.

559156 Rev. 8
Format Details
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FITC
Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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FITC
Blue 488 nm
494 nm
518 nm
559156 Rev.8
Citations & References
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Development References (9)

  1. Allman D, Li J, Hardy RR. Commitment to the B lymphoid lineage occurs before DH-JH recombination. J Exp Med. 1999; 189(4):735-740. (Biology). View Reference
  2. Allman D, Lindsley RC, DeMuth W, Rudd K, Shinton SA, Hardy RR. Resolution of three nonproliferative immature splenic B cell subsets reveals multiple selection points during peripheral B cell maturation. J Immunol. 2001; 167(12):6834-6840. (Biology). View Reference
  3. Auerbach R, Huang H, Lu L. Hematopoietic stem cells in the mouse embryonic yolk sac. Stem Cells. 1996; 14(3):269-280. (Biology). View Reference
  4. Jordan CT, McKearn JP, Lemischka IR. Cellular and developmental properties of fetal hematopoietic stem cells. Cell. 1990; 61(6):953-963. (Biology). View Reference
  5. Lacaud G, Carlsson L, Keller G. Identification of a fetal hematopoietic precursor with B cell, T cell, and macrophage potential. Immunity. 1998; 9(6):827-838. (Biology). View Reference
  6. Li YS, Wasserman R, Hayakawa K, Hardy RR. Identification of the earliest B lineage stage in mouse bone marrow. Immunity. 1996; 5(6):527-535. (Biology). View Reference
  7. McKearn JP, Baum C, Davie JM. Cell surface antigens expressed by subsets of pre-B cells and B cells. J Immunol. 1984; 132(1):332-339. (Immunogen). View Reference
  8. Paige CJ, Gisler RH, McKearn JP, Iscove NN. Differentiation of murine B cell precursors in agar culture. Frequency, surface marker analysis and requirements for growth of clonable pre-B cells. Eur J Immunol. 1984; 14(11):979-987. (Biology). View Reference
  9. Szilvassy SJ, Cory S. Phenotypic and functional characterization of competitive long-term repopulating hematopoietic stem cells enriched from 5-fluorouracil-treated murine marrow. Blood. 1993; 81(9):2310-2320. (Biology). View Reference
View All (9) View Less
559156 Rev. 8

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