AZDye 555 Tyramide (CCT-1540)

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Description

Orange fluorescent reagent widely used for tyramide signal amplification (TSA) in IHC, ICC, FISH and multicolor FISH. Tyramide signal amplification (TSA) has proven to be a particularly versatile and powerful enzyme amplification technique with improved assay sensitivity. TSA is based on the ability of HRP, in the presence of low concentrations of hydrogen peroxide, to convert labeled tyramine-containing substrate into an oxidized, highly reactive free radical that can covalently bind to tyrosine residues at or near the HRP.

AZDye™ 555 Tyramide (Alexa Fluor® 555 Tyramide equivalent) is a component of many tyramide signal amplification kits, including SuperBoost®. This reagent can be used with any antibody conjugated to HRP for tyramide signal amplification for detection of low abundance targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC ), and in situ hybridization (ISH).

Specifications

Unit Size150 slides, 1500 slides
Abs/Em Maxima555/572 nm
Extinction Coefficient
155,000
Flow Cytometry Laser Line
532 nm or 555 nm
Microscopy Laser Line
532 nm or 555 nm
Spectrally Similar DyesAlexa Fluor® 555, CF® 555, DyLight® 549, Cy3 Dye
Molecular weight966.16 (protonated)
CASN/A
SolubilityWater, DMSO, DMF
AppearanceRed solid
Storage Conditions-20°C. Desiccate
Shipping ConditionsAmbient temperature

Abs/Em Spectra

Af555

FAQs

150 slides has 0.24 µmol (depending on molecular weight this is ~200 µg)
1500 slides has 2.4 µmol (depending on molecular weight this is ~2 mg)

Here is a general protocol for the 150 slide unit size. To prepare 100x stock solution, we recommend reconstituting with 150 ul of DMSO. Storage once in solution: 100X stock solution in DMSO can be stored for 6 months at 2-8 °C protected from light. Avoid moisture.
Working solutions can be made fresh by adding 1 part 100X stock solution, 1 part 0.15% H202, and 100 part buffer.
For example, you can add 5 µL of 100X Tyramide stock solution and 5 µL 0.15% H2O2 to 500 µL of buffer. The final concentration of H2O2 is 0.0015%.
Typical reaction time following the HRP reagent is 2-10 minutes but should be optimized for each workflow. Following the TSA working solution incubation, wash thoroughly.

Applicable patents and legal notices are available at legal notices.

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