Multiparameter flow cytometric analysis of CD22 expression on Human peripheral blood leucocyte populations. Human whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220). The leucocytes were then stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or BD Horizon™ RB780 Mouse Anti-Human CD22 antibody (Cat. No. 569354/569355; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD22 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter signals of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
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Multiparameter flow cytometric analysis of CD22 expression on Human peripheral blood leucocyte populations. Human whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220). The leucocytes were then stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or BD Horizon™ RB780 Mouse Anti-Human CD22 antibody (Cat. No. 569354/569355; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD22 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter signals of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Product Details
BD Horizon™
BL-CAM; Siglec-2; Bgp135; Lyb8; LPAP
Human (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl/test
V CD22.14
933
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 to the dye under optimum conditions and unreacted dye was removed.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® 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 BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Human BD Fc Block™ RUO
Size
0.25 mg
Cat No.
564220
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
RB780 Mouse IgG1, κ Isotype Control RUO
Size
50 µg
Cat No.
568532
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569355 Rev. 2
Antibody Details
HIB22
The HIB22 monoclonal antibody specifically binds to CD22. CD22 is a 130-140 kDa glycosylated type I integral membrane protein present on the surface of mature B cells. CD22 is expressed in the cytoplasm of virtually all B cells except plasma cells. CD45RO antigen on T cells and CD75 antigen on B cells have been identified as ligands for CD22. CD22 has been reported to participate in B-cell activation and also as an adhesion molecule. Although the immunobiology of this antigen has not been fully elucidated, reports indicate that ligation of CD22 induces constitutive internalization of the molecule followed by complete degradation.
569355 Rev. 2
Format Details
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
Blue 488 nm
498 nm
781 nm
569355 Rev.2
Citations & References
View product citations for antibody "569355" on CiteAb
Development References (5)
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Clark EA. CD22, a B cell-specific receptor, mediates adhesion and signal transduction. J Immunol. 1993; 150(11):4715-4718. (Biology). View Reference
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Kehrl JH. CD22 Workshop Panel report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:523-525.
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
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Shan D, Press OW. Constitutive endocytosis and degradation of CD22 by human B cells. J Immunol. 1995; 154(9):4466-4475. (Biology). View Reference
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Stamenkovic I, Sgroi D, Aruffo A, Sy MS, Anderson T. The B lymphocyte adhesion molecule CD22 interacts with leukocyte common antigen CD45RO on T cells and alpha 2-6 sialyltransferase, CD75, on B cells. Cell. 1991; 66(6):1133-1144. (Biology). View Reference
569355 Rev. 2
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.