Skip to main content Skip to navigation
BB515 Mouse Anti-Human CD2
BB515 Mouse Anti-Human CD2
Flow cytometric analysis of CD2 expression on human peripheral blood lymphocytes- Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Whole blood was stained with either BD Horizon BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human CD2 antibody (Cat. No. 565264; bold solid line histograms). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Alternatively, cells were stained with FITC Anti-Human CD2 antibody (Cat. No. 555326/556608/561759; thin solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202).        Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD2 antibody versus its Ig Isotype Control (Left Panel), and BB515 Anti-CD2 antibody versus FITC Anti-CD2 antibody (Right Panel). The fluorescence histograms showing CD2 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD2 expression on human peripheral blood lymphocytes- Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Whole blood was stained with either BD Horizon BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human CD2 antibody (Cat. No. 565264; bold solid line histograms). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Alternatively, cells were stained with FITC Anti-Human CD2 antibody (Cat. No. 555326/556608/561759; thin solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202).        Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD2 antibody versus its Ig Isotype Control (Left Panel), and BB515 Anti-CD2 antibody versus FITC Anti-CD2 antibody (Right Panel). The fluorescence histograms showing CD2 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
Down Arrow Up Arrow


BD Horizon™
LFA-2; LFA-3 receptor; SRBC; T-cell surface antigen T11/Leu-5; erythrocyte receptor
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Human Phytohemagglutinin-treated Lymphoblasts
Flow cytometry (Routinely Tested)
5 µl
IV T085; VI 6T-078
914
AB_2739139
Aqueous buffered solution containing ≤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 BD Horizon™ BB515 under optimum conditions and unconjugated antibody was removed.

Recommended Assay Procedures

  BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  5. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565264 Rev. 3
Antibody Details
Down Arrow Up Arrow
RPA-2.10

  The RPA-2.10 monoclonal antibody specifically binds to CD2 which is also known as Lymphocyte-function antigen-2 (LFA-2), LFA-3 receptor, Erythrocyte receptor, Sheep red blood cell (SRBC) receptor, or T-cell surface antigen T11/Leu-5. CD2 is a 50 kDa type I transmembrane glycoprotein. CD2 belongs to the immunoglobulin superfamily of proteins along with its primary ligand, LFA-3 (CD58). It is expressed on the surface of ~80-90% of human peripheral blood lymphocytes, greater than 95% of thymocytes, all T lymphocytes that form E-rosettes, and a subset of NK cells. CD2 functions as an adhesion receptor that binds to CD58 resulting in the activation of CD2-positive T cells and NK cells and in the regulation of their cytolytic activities.

The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

565264 Rev. 3
Format Details
Down Arrow Up Arrow
BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BB515
Blue 488 nm
490 nm
515 nm
565264 Rev.3
Citations & References
Down Arrow Up Arrow
View product citations for antibody "565264" on CiteAb

Development References (7)

  1. Aversa GG, Bishop GA, Suranyi MG, Hall BM. RPA-2.10: an anti-CD2 monoclonal antibody that inhibits alloimmune responses and monitors T cell activation. Transplant Proc. 1987; 19(1):277-278. (Immunogen: Flow cytometry, Immunoprecipitation, Inhibition). View Reference
  2. Hahn WC, Burakoff SJ, Bierer BE. Signal transduction pathways involved in T cell receptor-induced regulation of CD2 avidity for CD58. J Immunol. 1993; 150(7):2607-2619. (Biology). View Reference
  3. Jonker M, Slingerland W. Reactivity of mAb specific for human CD markers with Rhesus monkey leucocytes. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1058-1063.
  4. Kato K. CD2 Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:39-43.
  5. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  6. Sopper S, Stahl-Hennig C, Demuth M, Johnston IC, Dorries R, ter Meulen V. Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys. Cytometry. 1997; 29(4):351-362. (Clone-specific: Flow cytometry). View Reference
  7. Suranyi MG, Bishop GA, Clayberger C, et al. Lymphocyte adhesion molecules in T cell-mediated lysis of human kidney cells. Kidney Int. 1991; 39(2):312-319. (Clone-specific: Flow cytometry, Inhibition). View Reference
View All (7) View Less
565264 Rev. 3

 

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.