DyLight^TM Fluorescent Conjugates
KPL’s DyLight conjugates offer a brilliant choice in a variety of immunofluorescence detection applications. They combine the sensitivity and reproducibility of our affinity purified secondary antibodies with a series of DyLight dyes that span the light spectra from visible to infrared.  They are brighter than fluorescein, rhodamine, Cy™3 and Cy5 and offer comparable brightness and photostability to Alexa conjugates. They are ideal for use in the following applications:

• Fluorescent Western blotting
• Fluorescence microscopy
• Flow cytometry
• FLISA (plate or surface-based functional assays)
• Microarray
• High content imaging

Feature	Benefit
Intense fluorescence	High sensitivity, bright signal, conjugate conservation
Excellent photostability	More photostable than Cy, FITC, comparable to Alexa
Sensitive, specific antibodies	High signal to noise ratio, low background
Consistent conjugate performance	No need to change assay parameters from lot-to-lot
Wide conjugate selection	DyLight conjugates to eleven animal species and streptavidin; specificities to whole antibody, heavy chain (gamma and mu) and F(ab')2 fragment
Well-differentiated emission spectra without overlapping between fluorophore channels	Enables specific, multi-color analysis
pH insensitive over range 4 - 9	Conjugates compatible with many buffers.
Instrument compatibility	Ideal for use with all common fluorescence instrumentation.

Excitation, Absorption and Emission Spectra for KPL DyLight Dyes

DyLight 405	DyLight 488	DyLight 549	DyLight 594	DyLight 633	DyLight 649	DyLight 680	DyLight 800
Excitation	Excitation	Excitation	Excitation	Excitation	Excitation	Excitation
Absorption	Absorption	Absorption	Absorption	Absorption	Absorption	Absorption	Absorption
Emission	Emission	Emission	Emission	Emission	Emission	Emission	Emission

 

DyLight Dyes Emission Spectra

DyLight Dyes
KPL offers eight of the most practical DyLight conjugates with well differentiated excitation and emission spectra.  Their spectral separation facilitates the design of multiple labeling experiments in fluorescence microscopy, flow cytometry, Western blotting and microwell plate assays.

DyLight 405

The Antibodies
Our extensive line of DyLight conjugates is available across a range of animal species, including human, mouse, rabbit and rat antibodies, as well as other animal species and streptavidin. They are affinity purified and in some cases further adsorbed to minimize cross-reactivity between animal species or shared reactivity with other immunoglobulin classes. KPL mouse antibodies are cross-adsorbed against human serum to minimize cross-reactivity. Anti-heavy and light chain antibodies provide species-specific detection of the IgG class. Antibodies directed against heavy chains react specifically with the heavy chain in the product name; light chain reactivity has been removed. F(ab’)₂ fragment antibodies are also available for assays requiring extremely low background levels and absence of F(c)-mediated binding.

DyLight Offers Performance Comparable to Alexa

In a plate-based FLISA KPL's DyLight 649 rabbit anti-goat IgG (H+L) conjugate and Alexa 647 rabbit anti-goat conjugate were added to a serially diluted goat IgG plate. The data demonstrate that the DyLight conjugate produces fluorescense signal comparable to that produced by the Alexa conjugate.

Excellent Reproducibility Reduces Need to Re-Optimize

Lot-to-lot comparisons provide an indicator of conjugate consistency with respect to both antibody performance and fluorescence intensity. KPL carefully controls its antibody production and fluorochrome conjugation processes to achieve the highest possible quality and consistency as demonstrated by the results in the figure below.

Three production lots of Goat Anti-human IgG (H+L) were labeled with DyLight 488. An ELISA was performed by adding 0.10 mL of a 10,000 ng/mL solution to a serially diluted human IgG microwell plate with a starting concentration of 10,000 ng/mL.  Variability among lots was extremely low (less than 4% Relative Standard Deviation).