BD Horizon™ RB613 Streptavidin
Multiparameter flow cytometric analyses using BD Horizon™ RB613 Streptavidin Left Plot: CD45R/B220 expression on viable Mouse splenic leukocytes. Mouse splenocytes were either labeled with Biotin Rat IgG2a κ Isotype Control (Cat. No. 553928; dashed line histogram) or with Biotin Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553086; solid line histogram). The cells were then washed and secondarily stained with BD Horizon™ RB613 Streptavidin (Cat. No. 571111) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The histogram showing CD45R/B220 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic leukocytes. Right Plot: CD3 expression on Human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were either labeled with Biotin Mouse IgG1, κ Isotype Control (Cat. No. 569597; dashed line histogram) or with Biotin Mouse Anti-Human CD3 antibody (solid line histogram; Cat. No. 555331). The cells were then washed and secondarily stained with BD Horizon™ RB613 Streptavidin (Cat. No. 571111) at 0.25 µg/test. The histogram showing CD3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lympocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analyses using BD Horizon™ RB613 Streptavidin Left Plot: CD45R/B220 expression on viable Mouse splenic leukocytes. Mouse splenocytes were either labeled with Biotin Rat IgG2a κ Isotype Control (Cat. No. 553928; dashed line histogram) or with Biotin Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553086; solid line histogram). The cells were then washed and secondarily stained with BD Horizon™ RB613 Streptavidin (Cat. No. 571111) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The histogram showing CD45R/B220 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic leukocytes. Right Plot: CD3 expression on Human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were either labeled with Biotin Mouse IgG1, κ Isotype Control (Cat. No. 569597; dashed line histogram) or with Biotin Mouse Anti-Human CD3 antibody (solid line histogram; Cat. No. 555331). The cells were then washed and secondarily stained with BD Horizon™ RB613 Streptavidin (Cat. No. 571111) at 0.25 µg/test. The histogram showing CD3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lympocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Horizon™
Flow cytometry (Routinely Tested)
0.1 mg/ml
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. Streptavidin was conjugated with dye under optimum conditions, and unconjugated Streptavidin and free dye were removed
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.
- 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).
- CF™ is a trademark of Biotium, Inc.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Biotin Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
569597
Biotin Rat IgG2a κ Isotype Control RUO
Size
0.25 mg
Cat No.
553928
Biotin Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
555331
Biotin Rat Anti-Mouse CD45R/B220 RUO
Size
0.5 mg
Cat No.
553086
571111 Rev. 1
Antibody Details
Streptavidin is a non-glycosylated protein that is prepared chromatographically from the bacterium Streptomyces avidinii. Streptavidin homotetramers have a particularly high, non-covalent binding affinity for biotin. When conjugated with fluorochromes, streptavidin has been widely used with biotin-conjugated antibodies and other biotinylated specific-binding molecules (eg, recombinant proteins and lectins) to stain cells and tissues for subsequent multiparameter analysis by flow cytometry, fluorescence microscopy and imaging. Likewise, when conjugated with an enzyme (eg, Horseradish Peroxidase or Alkaline Phosphatase) and coupled with a colorimetric or luminescent substrate development system, streptavidin has found widespread use along with biotinylated antibodies in a number of applications including Western blot, ELISA, ELISPOT, immunocytochemistry and immunohistochemistry.
571111 Rev. 1
Format Details
RB613
The BD Horizon RealBlue™ 613 (RB613) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492-nm and an emission maximum (Em Max) at 613-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB613 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with reduced excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB613 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-CF594. RB613 is on average brighter than PE-CF594 off the blue laser.
RB613
492 nm
613 nm
571111 Rev.1
Citations & References
View product citations for antibody "571111" on CiteAb
Development References (2)
-
Diamandis EP, Christopoulos TK. The biotin-(strept)avidin system: principles and applications in biotechnology. Clin Chem. 1991; 37(5):625-636. (Methodology). View Reference
-
Shapiro HM. Practical flow cytometry, 4th ed.. Hoboken, N.J.: Wiley-Liss; 2003:1-681.
571111 Rev. 1
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.