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BB515 Mouse Anti-Human CD235ab (Glycophorin A/B)
565233Image1.png

Flow cytometric analysis of CD235ab (Glycophorin A/B) expression on Human peripheral blood erythrocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies.  Human whole blood was stained with either BD Horizon BB515 Mouse IgG2b, κ Isotype Control (Cat. No. 564510; dashed line histogram) or BD Horizon™ BB515 Mouse Anti-Human CD235ab (Glycophorin A/B) antibody (Cat. No. 565233/565234; bold solid line histogram). Alternatively, the cells were stained with FITC Anti-Human CD235ab (Glycophorin A/B) antibody (Cat. No. 559943/561017; thin solid line histogram).  Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD235ab (Glycophorin A/B) antibody versus its Ig Isotype Control (Left Plot), and BB515 Anti-CD235ab (Glycophorin A/B) antibody versus FITC Anti-CD235ab (Glycophorin A/B) antibody (Right Plot). The fluorescence histograms showing CD235ab (Glycophorin A/B) expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact erythrocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

565233_565234-2a.png

Flow cytometric analysis of CD235ab (Glycophorin A/B) expression on Human peripheral blood erythrocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies.  Human whole blood was stained with either BD Horizon BB515 Mouse IgG2b, κ Isotype Control (Cat. No. 564510; dashed line histogram) or BD Horizon™ BB515 Mouse Anti-Human CD235ab (Glycophorin A/B) antibody (Cat. No. 565233/565234; bold solid line histogram). Alternatively, the cells were stained with FITC Anti-Human CD235ab (Glycophorin A/B) antibody (Cat. No. 559943/561017; thin solid line histogram).  Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD235ab (Glycophorin A/B) antibody versus its Ig Isotype Control (Left Plot), and BB515 Anti-CD235ab (Glycophorin A/B) antibody versus FITC Anti-CD235ab (Glycophorin A/B) antibody (Right Plot). The fluorescence histograms showing CD235ab (Glycophorin A/B) expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact erythrocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

565233Image1.png 565233_565234-2a.png hCD235a_BB515_vs_FITC.png

Flow cytometric analysis of CD235ab (Glycophorin A/B) expression on Human peripheral blood erythrocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies.  Human whole blood was stained with either BD Horizon BB515 Mouse IgG2b, κ Isotype Control (Cat. No. 564510; dashed line histogram) or BD Horizon™ BB515 Mouse Anti-Human CD235ab (Glycophorin A/B) antibody (Cat. No. 565233/565234; bold solid line histogram). Alternatively, the cells were stained with FITC Anti-Human CD235ab (Glycophorin A/B) antibody (Cat. No. 559943/561017; thin solid line histogram).  Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD235ab (Glycophorin A/B) antibody versus its Ig Isotype Control (Left Plot), and BB515 Anti-CD235ab (Glycophorin A/B) antibody versus FITC Anti-CD235ab (Glycophorin A/B) antibody (Right Plot). The fluorescence histograms showing CD235ab (Glycophorin A/B) expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact erythrocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Flow cytometric analysis of CD235ab (Glycophorin A/B) expression on Human peripheral blood erythrocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies.  Human whole blood was stained with either BD Horizon BB515 Mouse IgG2b, κ Isotype Control (Cat. No. 564510; dashed line histogram) or BD Horizon™ BB515 Mouse Anti-Human CD235ab (Glycophorin A/B) antibody (Cat. No. 565233/565234; bold solid line histogram). Alternatively, the cells were stained with FITC Anti-Human CD235ab (Glycophorin A/B) antibody (Cat. No. 559943/561017; thin solid line histogram).  Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD235ab (Glycophorin A/B) antibody versus its Ig Isotype Control (Left Plot), and BB515 Anti-CD235ab (Glycophorin A/B) antibody versus FITC Anti-CD235ab (Glycophorin A/B) antibody (Right Plot). The fluorescence histograms showing CD235ab (Glycophorin A/B) expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact erythrocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Product Details
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BD Horizon™
CD235a; CD235b; Glycophorin-A; Glycophorin-B; GYPA; GYPB; GPA; GPB
Human (QC Testing)
Mouse IgG2b, κ
Flow cytometry (Routinely Tested)
0.2 mg/ml
VII 70299
2993, 2994
AB_2744331
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

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 unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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Recommended Assay Procedures

Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended application.

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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.

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Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  8. For U.S. patents that may apply, see bd.com/patents.
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565234 Rev. 3
Antibody Details
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GA-R2 (HIR2)

The GA-R2 (also known as HIR2) monoclonal antibody specifically binds to CD235a and CD235b. CD235a is also known as Glycophorin A (GYPA, GPA, GLPA), Sialoglycoprotein alpha, MN sialoglycoprotein, or PAS-2. CD235b is otherwise known as Glycophorin B (GYPB, GPB, GLPB), Sialoglycoprotein delta, SS-active sialoglycoprotein, or PAS-3. CD235a and CD235b are type I transmembrane sialoglycoproteins that are expressed on human erythrocytes, erythroid precursor cells and certain leukemic cell types. CD235a carries blood group M and N antigens, whereas CD235b contains S, s, and U antigens. This antibody is useful for the identification and characterization of erythrocytes, certain myeloid leukemic cell types, and studies of erythroid cell development and infectious diseases with erythrocyte involvement. Glycophorins may play a role in preventing cell agglutination.

565234 Rev. 3
Format Details
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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.
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BB515
490 nm
515 nm
565234 Rev.3
Citations & References
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View product citations for antibody "565234" on CiteAb

Development References (8)

  1. Bain BJ. Leukemia diagnosis: A guide to the FAB classification. 1990.
  2. Blanchard D, Roux YP-L, Vusio P, Follea G. Characterization of monoclonal antibodies directed to human red blood cell glycophorins A and B. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:579-582.
  3. Gross S, Helm K, Gruntmeir JJ, Stillman WS, Pyatt DW, Irons RD. Characterization and phenotypic analysis of differentiating CD34+ human bone marrow cells in liquid culture. Br J Haematol. 1997; 5(318):326. (Clone-specific: Flow cytometry). View Reference
  4. Keren DF, Hanson CA, Hurtubise PE. David F. Keren, Curtis A. Hanson, Paul E. Hurtubise., ed. Flow cytometry and clinical diagnosis. Chicago: ASCP Press; 1994:1-676.
  5. Loken MR, Civin CI, Bigbee WL, Langlois RG, Jensen RH. Coordinate glycosylation and cell surface expression of glycophorin A during normal human erythropoiesis. Blood. 1987; 70(6):1959-1961. (Biology). View Reference
  6. Nakahata T, Okumura N. Cell surface antigen expression in human erythroid progenitors: erythroid and megakaryocytic markers. Leuk Lymphoma. 1994; 13(5-6):401-409. (Biology). View Reference
  7. Rogers CE, Bradley MS, Palsson BO, Koller MR. Flow cytometric analysis of human bone marrow perfusion cultures: erythroid development and relationship with burst-forming units-erythroid. Exp Hematol. 1996; 24(5):597-604. (Biology). View Reference
  8. Zola H, Swart B, Boumsell L, Mason DY. Human Leucocyte Differentiation Antigen nomenclature: update on CD nomenclature. Report of IUIS/WHO Subcommittee.. J Immunol Methods. 2003; 275(1-2):1-8. (Clone-specific: Flow cytometry). View Reference
View All (8) View Less
565234 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.

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.