BD Horizon™ RB780 Mouse Anti-Biotin
Clone BK-1/39 (RUO)
Multiparameter flow cytometric analysis of CD25 expression on viable Mouse splenic lymphocytes. Mouse splenic leukocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) and stained with Biotin Rat Anti-Mouse CD25 antibody (Cat. No. 567831). The cells were washed and then secondarily stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or BD Horizon™ RB780 Mouse Anti-Biotin antibody (Cat. No. 570944/571016; Right Plot) at 0.125 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing CD25 expression (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD19 expression on viable Mouse splenic lymphocytes. Left Panel - One-layer staining: Mouse splenic leukocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142] and then stained with either BD Horizon™ RB780 Rat IgG2a, κ Isotype Control (Cat. No. 568698; dashed line histogram) or BD OptiBuild™ RB780 Rat Anti-Mouse CD19 antibody (Cat. No. 755522; solid line histogram). Right Panel - Two-layer staining: Mouse splenic leukocytes were preincubated with Mouse BD Fc Block™ and then stained with Biotin Rat Anti-Mouse CD19 antibody (Cat. No. 553784), washed, and secondarily stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Biotin antibody (Cat. No. 570944/571016; solid histogram) at 0.125 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing CD19 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
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Multiparameter flow cytometric analysis of CD25 expression on viable Mouse splenic lymphocytes. Mouse splenic leukocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) and stained with Biotin Rat Anti-Mouse CD25 antibody (Cat. No. 567831). The cells were washed and then secondarily stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or BD Horizon™ RB780 Mouse Anti-Biotin antibody (Cat. No. 570944/571016; Right Plot) at 0.125 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing CD25 expression (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD19 expression on viable Mouse splenic lymphocytes. Left Panel - One-layer staining: Mouse splenic leukocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142] and then stained with either BD Horizon™ RB780 Rat IgG2a, κ Isotype Control (Cat. No. 568698; dashed line histogram) or BD OptiBuild™ RB780 Rat Anti-Mouse CD19 antibody (Cat. No. 755522; solid line histogram). Right Panel - Two-layer staining: Mouse splenic leukocytes were preincubated with Mouse BD Fc Block™ and then stained with Biotin Rat Anti-Mouse CD19 antibody (Cat. No. 553784), washed, and secondarily stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Biotin antibody (Cat. No. 570944/571016; solid histogram) at 0.125 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing CD19 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Horizon™
Mouse IgG1, κ
Biotin-conjugated Keyhole Limpet Hemocyanin (KLH)
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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.
- For U.S. patents that may apply, see bd.com/patents.
- 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 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.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
Companion Products
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RB780 Mouse IgG1, κ Isotype Control RUO
Size
50 µg
Cat No.
568532
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Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
RB780 Rat IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
568698
RB780 Rat Anti-Mouse CD19 RUO
Size
50 µg
Cat No.
755522
Biotin Rat Anti-Mouse CD19 RUO
Size
0.5 mg
Cat No.
553784
570944 Rev. 1
Antibody Details
BK-1/39
The BK-1/39 monoclonal antibody specifically recognizes biotin. Biotin is a water-soluble B complex vitamin that functions as a coenzyme which is required for the cellular metabolism of proteins and fats and the production of fatty acids. Although required by all organisms, biotin synthesis is limited to bacteria, yeasts, molds, algae, and some plants. Biotin is also useful for tagging molecules such as nucleic acids or proteins including antibodies. The BK-1/39 antibody can used to detect biotinylated target molecules and antibodies especially when signal amplification is desired. Fluorescent BK-1/39 antibody conjugates can provide a useful alternative to fluorescent avidin conjugates in order to minimize background staining and maximize signal intensity.
570944 Rev. 1
Format Details
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
Blue 488 nm
498 nm
781 nm
570944 Rev.1
Citations & References
View product citations for antibody "570944" on CiteAb
Development References (4)
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Levit-Zerdoun E, Becker M, Pohlmeyer R, et al. Survival of Igα-Deficient Mature B Cells Requires BAFF-R Function.. J Immunol. 2016; 196(5):2348-60. (Clone-specific: Flow cytometry). View Reference
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Mavrangelos C, Swart B, Nobbs S, Nicholson IC, Macardle PJ, Zola H. Detection of low-abundance membrane markers by immunofluorescence--a comparison of alternative high-sensitivity methods and reagents.. J Immunol Methods. 2004; 289(1-2):169-78. (Methodology: Flow cytometry). View Reference
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Pishesha N, Bilate AM, Wibowo MC, et al. Engineered erythrocytes covalently linked to antigenic peptides can protect against autoimmune disease.. Proc Natl Acad Sci U S A. 2017; 114(12):3157-3162. (Clone-specific: Flow cytometry). View Reference
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Zempleni J, Wijeratne SS, Hassan YI. Biotin.. Biofactors. 2009; 35(1):36-46. (Biology). View Reference
570944 Rev. 1
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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.