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.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The 15D3 monoclonal antibody specifically recognizes P-glycoprotein which is also known as CD243, ATP-binding cassette subfamily B member 1 (ABCB1) or Multidrug resistance protein 1 (MDR1). P-glycoprotein is a ~170 kDa transmembrane glycoprotein protein that spans the membrane 12 times. P-glycoprotein acts as an ATP-dependent efflux pump for a large variety of drugs. It may be expressed at high levels by multidrug resistant (MDR) tumor cells. This efflux activity may lead to cellular resistance to the drugs used in chemotherapy. P-glycoprotein is present in many normal cell types including endothelial, epithelial, or secretory cells, and might protect them from naturally occurring xenobiotics.
Development References (9)
-
Beck WT, Grogan TM, Willman CL, et al. Methods to detect P-glycoprotein-associated multidrug resistance in patients' tumors: consensus recommendations.. Cancer Res. 1996; 56(13):3010-20. (Biology). View Reference
-
Chan HS, Haddad G, Zheng L, Bradley G, Dalton WS, Ling V. Sensitive immunofluorescence detection of the expression of P-glycoprotein in malignant cells.. Cytometry. 1997; 29(1):65-75. (Biology). View Reference
-
Chaudhary PM, Mechetner EB, Roninson IB. Expression and activity of the multidrug resistance P-glycoprotein in human peripheral blood lymphocytes.. Blood. 1992; 80(11):2735-9. (Biology). View Reference
-
Dalton WS, Durie BG, Alberts DS, Gerlach JH, Cress AE. Characterization of a new drug-resistant human myeloma cell line that expresses P-glycoprotein.. Cancer Res. 1986; 46(10):5125-30. (Biology). View Reference
-
Drach D, Zhao S, Drach J, et al. Subpopulations of normal peripheral blood and bone marrow cells express a functional multidrug resistant phenotype.. Blood. 1992; 80(11):2729-34. (Biology). View Reference
-
Filipits M, Suchomel RW, Dekan G, et al. MRP and MDR1 gene expression in primary breast carcinomas.. Clin Cancer Res. 1996; 2(7):1231-7. (Biology). View Reference
-
Klimecki WT, Futscher BW, Grogan TM, Dalton WS. P-glycoprotein expression and function in circulating blood cells from normal volunteers.. Blood. 1994; 83(9):2451-8. (Biology). View Reference
-
List AF. Role of multidrug resistance and its pharmacological modulation in acute myeloid leukemia.. Leukemia. 1996; 10(6):937-42. (Biology). View Reference
-
Shi T, Wrin J, Reeder J, Liu D, Ring DB. High-affinity monoclonal antibodies against P-glycoprotein. Clin Immunol Immunopathol. 1995; 76(1):44-51. (Immunogen: ELISA, Flow cytometry, Immunoprecipitation). View Reference
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.