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Streptavidin and Avidin are proteins that bind to biotin with high affinity. Streptavidin conjugates with fluorophores or enzymes are utilized as a secondary labeling method to detect biotinylated molecules in common research applications such as: • flow cytometry • tetramer staining • immunofluorescence microscopy • ELISA • in situ hybridization • immunohistochemistry • microarrays • western blot, and other applications.
BioLegend provides a wide array of streptavidin conjugates, including a conjugate with the new Brilliant Violet 421™, a novel fluorophore excited by the violet laser giving unmatched performance in flow cytometry and microscopy.
Immortalized mouse bone marrow-derived macrophages, treated with LPS (100 ng/ml for 24 h), were stained with Alexa Fluor® 594 Wheat Germ Agglutinin (red) and biotin anti-mouse IL-10 (clone JES5-16E3), followed by Brilliant Violet 421™ Streptavidin (light blue). Image was acquired on an Olympus FV1000 laser scanning confocal microscope at 60x lens, 2x zoom. Data provided by James Harris, Trinity College, Dublin.
Biotin CD56 + Brilliant Violet 421™ Streptavidin
Human NK cell line NK92 fixed and stained with biotin anti-CD56 (clone HCD56) and Brilliant Violet 421™Streptavidin (blue), phalloidin Alexa Fluor® 568 (red), and anti-perforin FITC (green) imaged on Olympus IX81 spinning disk confocal scope on 100 X objective NA 1.45. The first image is the DIC view (far left). The final image (far right) shows the merged fluorescence of the three stains. Data provided by Emily Mace and Jordan Orange, Children's Hospital of Philadelphia.
Biotin anti-mouse CD272 (BTLA) +
PE Streptavidin
C57BL/6 splenocytes stained with biotin anti-mouse CD272 (clone 8F4)(left) or biotin mouse IgG1 isotype control (right), followed by PE Streptavidin and CD3e (clone 145-2C11) APC.
HLA-A2/peptide Tetramer + PE Streptavidin or Brilliant Violet 421™ Streptavidin
Human PBMCs were stained with CD8 PerCP and labeled with NLVPMVATV-loaded human HLA-A2 tetramer bound to PE Streptavidin, Brilliant Violet 421™Streptavidin, or Pacific Blue™ Streptavidin. The data shows positive staining with a wide range of tetramer dilutions for PE and BV421™. Data provided by Rick Willis and John Altman, Emory/Yerkes.
