Description
The JC159 monoclonal antibody specifically binds to CD235a which is encoded by GYPA [glycophorin A (MNS blood group)] and known as Sialoglycoprotein alpha or MN sialoglycoprotein. This heavily glycosylated type I transmembrane protein forms a ~36 kDa homodimer that bears the blood group M and N antigens in its N-terminal extracellular region. The JC159 antibody reacts with CD235a (Glycophorin A) but does not crossreact with the homologous CD235b (Glycophorin B) molecule. In contrast, the GA-R2 (HIR2) antibody more broadly recognizes both Glycophorins A and B, ie, is specific for CD235ab (Glycophorin A/B). CD235a (Glycophorin A) is expressed on human erythrocytes, erythroid precursor cells and certain leukemic cell types. This highly expressed, negatively charged surface sialoglycoprotein contributes to the negative charge of the red cell membrane that may help prevent erythrocyte aggregation in the circulation. This glycophorin provides structural stability to the erythrocyte membrane by associating with other cell surface proteins such as the anion exchanger, SLC4A1 (Band 3). SLC4A1 mediates gas exchange—oxygen uptake and carbon dioxide release—across the erythrocyte membrane. CD235a (Glycophorin A) binds to multiple ligands including Siglec-5 (CD170) which may help mediate interactions between erythrocytes and immune cells, eg, during inflammation, immune surveillance, or removal of damaged or aged red blood cells. The malaria parasite Plasmodium falciparum binds to CD235a (Glycophorin A) by its surface Erythrocyte Binding Antigen 175 (EBA-175) which can promote parasite entry and infection of erythrocytes. Viruses including influenza A and B viruses, hepatitis A virus, Sendai virus, encephalomyocarditis virus (EMCV), and reoviruses similarly bind to this glycophorin which can facilitate viral infection of cells.
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. 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). Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
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 that minimize unconjugated dye and antibody.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
4. 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.
5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
8. An isotype control should be used at the same concentration as the antibody of interest.
9. 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.
