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BUV395 Mouse Anti-Human CD235a
BUV395 Mouse Anti-Human CD235a
Flow cytometric analysis of CD235a expression on human peripheral blood erythrocytes. Whole blood was stained with either BD Horizon™ BUV395 Mouse IgG2b, κ Isotype Control (Cat. No. 563558; dashed line histogram) or BD Horizon™ BUV395 Mouse Anti-Human CD235a antibody (Cat. No. 563810/565973; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of erythrocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD235a expression on human peripheral blood erythrocytes. Whole blood was stained with either BD Horizon™ BUV395 Mouse IgG2b, κ Isotype Control (Cat. No. 563558; dashed line histogram) or BD Horizon™ BUV395 Mouse Anti-Human CD235a antibody (Cat. No. 563810/565973; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of erythrocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
CD235ab; CD235a, Glycophorin-A, GYPA, GPA; CD235b; Glycophorin-B, GYPB, GPB
Human (QC Testing)
Mouse IgG2b, κ
Flow cytometry (Routinely Tested)
5 µl
VII 70299
AB_2738434
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 BUV395 under optimum conditions, and unconjugated antibody and free BD Horizon BUV395 were 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).

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 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.
565973 Rev. 2
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.

The antibody was conjugated to BD Horizon BUV395 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. With an Ex Max near 348 nm and an Em Max near 395 nm, BD Horizon BUV395 can be excited by the ultraviolet laser (355 nm) laser and detected with a 379/28 filter. This dye has been exclusively developed by BD Biosciences as an optimal dye for use on instruments equipped with the ultraviolet laser and has virtually no spillover into any other detector.

565973 Rev. 2
Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
Ultraviolet 355 nm
348 nm
395 nm
565973 Rev.2
Citations & References
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Development References (6)

  1. Bain BJ. Leukemia diagnosis: A guide to the FAB classification. 1990.
  2. 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
  3. 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.
  4. 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
  5. 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
  6. 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
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565973 Rev. 2

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