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AZDye™ 488 Biotin can be used for detecting and quantifying biotin binding sites of avidin, streptavidin or neutravidin. This reagent overcomes major shortcomings of commonly used Biotin-4-fluorescein – poor solubility in aqueous media and pH dependent absorbance and emission. AZDye™ 488 Biotin is a water soluble reagent and its green fluorescence is pH independent from pH 4 to pH 10. A flexible PEG3 spacer between biotin moiety and fluorescent tag minimize steric hindrance involved in binding to avidin, streptavidin or neutravidin.

AZDye™ 488 is a bright, and highly photostable, green-fluorescent probe optimally excited by the 488 nm laser line. This probe is water-soluble and its fluorescence is pH independent over a wide pH range. The brightness and photostability of blue dyes are best suited to direct imaging of low-abundance targets.

AZDye™ 488 is structurally identical to Alexa Fluor® 488. Its absorption/emission spectra is a perfect match to spectra of many other fluorescent dyes based on sulfonated rhodamine 110 core, including DyLight® 488, Alexa Fluor® 488, and CF® 488A.

DyLight® and Alexa Fluor® are a registered trademark of Thermo Fisher Scientific.

[caption id="attachment_25358" align="alignnone" width="1000"] Abs/Em Spectra[/caption]

Specifications

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AZDye 594 Biotin (Alexa Fluor® 594 Biotin equivalent) can be used for detecting and quantifying biotin binding sites of avidin, streptavidin or neutravidin. This reagent overcomes major shortcomings of commonly used Biotin-4-fluorescein – poor solubility in aqueous media and pH dependent absorbance and emission. AZDye 594 Biotin is a water soluble reagent and its red fluorescence is pH independent from pH 4 to pH 10. A flexible PEG3 spacer between biotin moiety and fluorescent tag minimize steric hindrance involved in binding to avidin, streptavidin or neutravidin.

[caption id="attachment_26156" align="alignnone" width="1000"] Abs/Em Spectra[/caption]

Specifications

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Trifunctional Click Chemistry Probes that incorporate a ligation handle, a biotin and a fluorophore have become a popular tool for for tandem labeling of proteins and subsequent detection or enrichment. However, due to the strong interaction between biotin and streptavidin harsh conditions are necessary for the elution of enriched proteins. This usually leads to contamination of the sample with non-specifically bound proteins and endogenously biotinlytated proteins, which complicates target identification.

Cleavable Trifunctional Click Chemistry Probes overcome this major drawback of the streptavidin-biotin affinity purification. These probes contain a biotin moiety linked to a “clickable” group and fluorescent dye through a spacer arm containing a cleavable Dde linker. Dde moiety is stable to rigorous, denaturing wash conditions, acidic or basic conditions including generally applied buffer systems to which the biological sample may be exposed. At the same time Dde linker can be quantitatively cleaved under mild aqueous buffered conditions with 2% hydrazine.

Another very important advantage of dual label probes over regular biotin probes is built-in control. Each step of enrichment process can easily followed either by UV-Vis (550 nm) or by more sensitive fluorescence spectroscopy. After elution form streptavidin beads target proteins containing TAMRA label can be easily distinguished from non-specifically bound proteins and endogenously biotinlytated proteins.

This new dual label probe should be useful to expand the new applications of standard biotin–streptavidin technology.

Specifications

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