BD Pharmingen™ Alexa Fluor™ 488 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™) [Cat. No. 553142] and stained with Biotin Rat Anti-Mouse CD25 antibody (Cat. No. 567831). The cells were washed and then secondarily stained with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 567121; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Biotin antibody (Cat. No. 570938/571010; Right Plot) at 0.25 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.
.png){kind=spacer}
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™) [Cat. No. 553142] and stained with Biotin Rat Anti-Mouse CD25 antibody (Cat. No. 567831). The cells were washed and then secondarily stained with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 567121; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Biotin antibody (Cat. No. 570938/571010; Right Plot) at 0.25 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.
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- 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.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
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.
Development References (4)
-
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
-
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
-
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
-
Zempleni J, Wijeratne SS, Hassan YI. Biotin.. Biofactors. 2009; 35(1):36-46. (Biology). 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.