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Preparation And Storage
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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
Companion Products
The L01.1 monoclonal antibody specifically binds to CD71 which is also known as the transferrin receptor (TFR). This type II transmembrane glycoprotein is expressed on cells as a disulfide-linked homodimer comprised of 95 kDa monomers. CD71 is expressed on activated lymphocytes, monocytes, macrophages, brain endothelium, and most proliferating cells. The transferrin receptor is also present on early erythroid cells but is lost as reticulocytes differentiate into mature erythrocytes. CD71 is lowly expressed on normal resting lymphocytes and is upregulated during lymphocyte responses to antigens or mitogens. The transferrin receptor is essential for iron transport into proliferating cells. Through an endocytic pathway, the transferrin receptor mediates cellular iron uptake by binding and internalizing iron that is bound to transferrin. After releasing iron within the low pH endosomal environment, transferrin and its receptor can be recycled to the cell surface.
Development References (7)
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Jefferies WA, Brandon MR, Hunt SV, Williams AF, Gatter KC, Mason DY. Transferrin receptor on endothelium of brain capillaries. Nature. 1998; 312(5990):162-163. (Biology). View Reference
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Judd W, Poodry CA, Strominger JL. Novel surface antigen expressed on dividing cells but absent from nondividing cells.. J Exp Med. 1980; 152(5):1430-5. (Biology). View Reference
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Larrick JW, Cresswell P. Modulation of cell surface iron transferrin receptors by cellular density and state of activation.. J Supramol Struct. 1979; 11(4):579-86. (Biology). View Reference
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Loken MR, Shah VO, Dattilio KL, Civin CI. Flow cytometric analysis of human bone marrow: I. Normal erythroid development.. Blood. 1987; 69(1):255-63. (Biology). View Reference
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Newman RA. Lymphoid receptors for transferrin.. Meth Enzymol. 1987; 150:723-45. (Biology). View Reference
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Phillips JH, Le AM, Lanier LL. Natural killer cells activated in a human mixed lymphocyte response culture identified by expression of Leu-11 and class II histocompatibility antigens.. J Exp Med. 1984; 159(4):993-1008. (Immunogen). View Reference
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Schwarting R, Stein H. Cluster report: CD71. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:455-460.
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.