Description
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.
Streptavidin was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.
Format
- FormatBB515
- Excitation SourceBlue 488 nm
- Excitation Max490nm
- Emission Max515nm
BB515 is a dye that was exclusively developed by BD Biosciences as a brighter alternative to FITC. This dye is up to seven times brighter than FITC and has less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously.
Other Formats→
- AKP
- APC
- APC-Cy™7
- APC-R700
- BUV395
- BUV496
- BUV563
- BUV615
- BUV661
- BUV737
- BUV805
- BV421
- BV510
- BV605
- BV650
- BV711
- BV786
- FITC
- HRP
- PE
- PE-CF594
- PE-Cy™5
- PE-Cy™7
- PerCP
- PerCP-Cy™5.5
- V450
- V500
Suggested Companion Products
Stain Buffer (BSA) RUO 500mLCat No: 554657
Stain Buffer (FBS) RUO 500mLCat No: 554656
Lysing Buffer RUO 100mLCat No: 555899
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) 2.4G2RUO 0.1mgCat No: 553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) 2.4G2RUO 0.5mgCat No: 553142
Biotin Rat Anti-Mouse CD45R/B220 RA3-6B2RUO 0.1mgCat No: 553085
Biotin Rat Anti-Mouse CD45R/B220 RA3-6B2RUO 0.5mgCat No: 553086
Resources & Tools | ||||||
---|---|---|---|---|---|---|
Spectrum Viewer | Panel Designer | SpectrumViewer | Download TDS | Download MSDS |
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
- 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.
- 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.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.