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02.03.2011  
Product Overview

NEW:
4-Color Imaging with DyLight 405 Conjugates

DyLight fluorescent dyes are a new family of dyes with improved brightness and photostability. The new DyLight 405-conjugated secondary antibodies are excited maximally at about 400 nm and fluoresce with a peak at about 421 nm (Figure 1). They are very bright and photostable when used with confocal microscopes equipped with a 405 nm laser and appropriate emission filter (Figure 2).

Under these conditions, it is possible to perform effective 4-color imaging with good color separation, good photostability, and high sensitivity in both aqueous and permanent mounting media. The combination of DyLight 405, DyLight 488, Rhodamine Red-X, and DyLight 649 provides for maximum color separation (Figure 3).

Other 4-color dye combinations, which may be equally effective but have slightly less color separation, include DyLight 405, DyLight 488, DyLight 549 (or Cy3), and DyLight 649 (Figure 4).

Read more and view Products
 
 
Peak wavelengths of absorption and emission maxima for DyLight-conjugated, affinity-purified secondary antibodies from Jackson ImmunoResearch.
 
  Figure 2. Mouse retinal tissue stained with DyLight 405 (pseudo-colored green)-donkey anti-rabbit IgG (JIR) with rabbit anti-GFAP (DAKO), DyLight 488 (pseudo-colored red) -donkey
Figure 1. Excitation (violet) and emission (blue) spectra for DyLight 405 conjugated to an affinity-purified secondary antibody from Jackson ImmunoResearch.   anti-mouse IgG (JIR) with mouse anti-neurofilament (Abcam), and DyLight 649 (pseudo-colored blue)-donkey anti-goat IgG (JIR) with goat anti-collagen IV (Chemicon).
 
  Figure 4. Mouse retinal tissue stained with DyLight 405 (pseudo-colored green)-donkey anti-rabbit IgG (JIR) with rabbit anti-GFAP (DAKO),DyLight 488 (pseudo-colored red)
Figure 3. Emission spectra of DyLight 405 (blue), DyLight 488 (green), Rhodamine Red-X (red), and DyLight 649 (brown) following conjugation to antibodies. The figure shows that effective 4-color imaging can be performed with maximum color separation using these dyes.   -donkey anti-mouse IgG (JIR) with mouse anti-neurofilament (Abcam), Cy3 (pseudo-colored blue)-donkey anti-chicken IgG (JIR) with chicken anti-vimentin (Chemicon), and DyLight 649 (pseudo-colored white)- donkey anti-goat IgG (JIR) with goat anti-collagen IV (Chemicon).

NEW:
Anti-Goat IgG, Light Chain Specific or Western Blotting after immunoprecipitation

Monoclonal Mouse Anti-Goat IgG, Light Chain Specific is a new addition to our family of anti-Light Chain specific antibodies for Western blotting after immunoprecipitation (IP) of antigens with primary antibodies. Anti-Light Chain specific antibodies react with native primary antibodies used for detecting specific protein bands on Western blots (Figure A). If diluted properly, anti-Light Chain specific antibodies do not bind to the reduced and denatured IgG heavy chain band (50kDa) on blots (Figure B,D, E, and F). Therefore by using anti-Light Chain specific antibodies , detection of antigens with molecular weights near 50 kDa is not obscured by large amounts of reduced and denatured IgG heavy chains from primary antibodies used for immunoprecipitation. View a complete listing of anti-light chain antibody conjugates

   
 
Detection of primary antibodies made in goat (against a 50kDa protein) by mouse anti-goat IgG, Light Chain (LC) specific (A). Heavy (50kDa) and light (25kDa) chains of reduced and SDS-denatured mouse IgG (B andC), goat IgG (D), rat IgG (E), and rabbit IgG (F) were separated by SDS-PAGE and detected on Western blots using HRP-goat anti-mouse IgG, LC specific (B), HRP-goat anti-mouse IgG (H+L) (C), HRP-mouse anti-goat IgG, LC specific (D), HRP-goat anti-rat IgG, LC specific (E), and HRP-mouse anti-rabbit IgG, LC specific (F), respectively. No heavy chain band was detected when anti-IgG, LC specific antibodies were used for detection (B, D, E, and F). However, both heavy and light chain bands were detected with anti-IgG (H+L) (C).
 
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