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Secondary Antibodies and Streptavidin

 

 

Introduction

 
Fluorescent secondary antibodies and streptavidin conjugates are used for the indirect detection of target antigens in many applications including fluorescent cell imaging, western blotting, immunohistochemistry and more. The advantages of using a fluorescently labeled secondary antibody and streptavidin conjugates include brighter signal, multiplexing capabilities, and ease of use. GeneCopoeia offers a wide selection of fluorescent dye conjugated secondary antibodies and streptavidin for your research, featuring our exceptionally bright and photostable Andy Fluor™ dyes.

Features

  • Bright fluorescence with low background: Similar performance with Alexa Fluor® conjugates
  • Excellent photostablility: Resistant to photobleaching
  • Great selection of fluorophores: Over 10 different Andy Fluor™ dyes for multicolor imaging
  • Instrument compatibility: Excitation and emission spectra fit to the most popular fluorescence instruments’ laser settings and emission filter sets.

 

Fluorescent Secondary Antibody and Streptavidin Selection Guide

Secondary Antibody Streptavidin
Andy Fluor™ Dyes Other Fluorescent Labels Biotin & HRP Labels Andy Fluor™ & Cy® Dyes
Andy Fluor™ 350 Cy 3 Biotin Andy Fluor™ 350
Andy Fluor™ 405M Cy 5 HRP Andy Fluor™ 488
Andy Fluor™ 430 Cy 5.5   Andy Fluor™ 555
Andy Fluor™ 488 Cy 7   Andy Fluor™ 594
Andy Fluor™ 555 FITC   Andy Fluor™ 647
Andy Fluor™ 568     Cy®3
Andy Fluor™ 594     Cy®5
Andy Fluor™ 610      
Andy Fluor™ 647      
Andy Fluor™ 680      
Andy Fluor™ 750      

Andy Fluor™ dye conjugates have brighter fluorescence than other fluorophores.

Figure 1. Comparison of relative fluorescence of goat anti-rabbit IgG antibody conjugates prepared from Andy Fluor™ 488, 555, and 647 with Cy®2, Cy®3, and Cy®5.

Figure 2. Flow cytometry comparison of the brightness of goat anti-mouse IgG antibody conjugates prepared from Andy Fluor™ 488, 555, and 647 with Cy®2, DyLight™ 488, Cy®3, DyLight™ 550, and DyLight™ 650.

Andy Fluor™ dye conjugates have better photostability than other fluorophores.


Figure 3. Comparison of the photobleaching rates of Andy Fluor™ 488 goat anti-mouse IgG (H+L) (L109B) with FITC goat anti-mouse IgG (H+L) (L146B). The cytoskeleton of HeLa cells was labeled with mouse monoclonal anti-α-tubulin antibody in combination with Andy Fluor™ 488 goat anti-mouse IgG (H+L) antibody (top series) or with mouse monoclonal anti-α-tubulin antibody in combination with FITC goat anti-mouse IgG (H+L) antibody (bottom series). The fluorescence imaging was taken at 60-second intervals (0, 60, and 120 seconds of exposure).

Figure 4. Immunofluorescent stain of α-tubulin in BEAS2BNNK cells. α-Tubulin in fixed and permeabilized BEAS2BNNK cells was labeled with anti-α-tubulin primary antibody, and then visualized with goat anti-mouse IgG antibodies conjugated with either Andy Fluor™ 488 (top, left), Andy Fluor™ 555 (top, right), Andy Fluor™ 568 (bottom, left), or Andy Fluor™ 647 (bottom, right). Nuclei are counterstained with DAPI (blue).

Figure 5. Immunofluorescent stain of CCSP in mouse lung tissue. FFPE samples of mouse lung were labeled with rabbit anti-CCSP primary antibody, and then visualized with green-fluorescent Andy Fluor™ 488 goat anti-rabbit IgG antibody (green). Nuclei were counterstained with DAPI (blue).

Figure 6. Immunofluorescent stain of CCSP in mouse lung tissue. FFPE samples of mouse lung were labeled with rabbit anti-CCSP primary antibody, and then visualized with either Andy Fluor™ 555 (red, left), or Andy Fluor™ 568 (red, right). Nuclei were counterstained with DAPI (blue).

Figure 7. Immunofluorescent stain of α-tubulin in BEAS2BNNK cells and CCSP in mouse lung tissue. α-Tubulin in fixed and permeabilized BEAS2BNNK cells was labeled with anti-α-tubulin primary antibody, followed by incubation with biotin goat anti-mouse IgG antibody, and then visualized with Andy Fluor™ 488 Streptavidin (green, left). FFPE samples of mouse lung were labeled with rabbit anti-CCSP primary antibody, followed by incubation with biotin goat anti-mouse IgG antibody, and then visualized with Andy Fluor™ 488 Streptavidin (green, middle and right). Nuclei were counterstained with DAPI (blue).