Description

For rapid, sensitive and accurate screening of potential Tyrosinase inhibitors

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Tyrosinase (EC 1.14.18.1) is a copper-binding enzyme that is expressed across a vast range of species ranging from bacteria and fungi to mammals. It is involved in two sequential reactions of the melanin synthesis pathway: first being the hydroxylation of a monophenol and second the conversion of an ortho-diphenol to a quinone. Quinone then undergoes a series of reactions including polymerization to form melanin.

Tyrosinase is of great interest to the agriculture industry since it causes browning of fruits, vegetable, and mushrooms, as well as to the cosmetic industry as it causes skin darkening. Development and screening of tyrosinase inhibitors, therefore, is very useful for conditions such as hyperpigmentation and melasma. Tyrosinase activity is significantly increased in melanoma. Therefore, the detection of tyrosinase activity could be promising as a specific diagnostic test for melanoma and may be useful in monitoring patient response to melanoma treatments.

This Tyrosinase Activity Assay Kit is a simple one-step, plate-based assay for the measurement of tyrosinase activity in biological samples. In this assay, tyrosinase catalyzes the conversion of a phenolic substrate to a Quinone intermediate, which reacts with the tyrosine enhancer forming a highly stable chromophore with absorbance at 520 nm. The assay can detect as low as 30 μU Tyrosinase in biological samples.

Highly reproducibility
Highly Sensitive Assay to Screen for Tyrosinase Activity
Stable formulation of ready to use Reaction Facilitator (tyrosinase)

PEG3-(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane)-(Amino-Tri-(endo-BCN-PEG2-ethoxymethyl)-methane) is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The DBCO groups are commonly used for copper-free Click Chemistry reactions due to their strain promoted high energy. The hydrophilic PEG chain allows for increased water solubility.

PEG3-(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane)-(Amino-Tri-(endo-BCN-PEG2-ethoxymethyl)-methane) is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The DBCO groups are commonly used for copper-free Click Chemistry reactions due to their strain promoted high energy. The hydrophilic PEG chain allows for increased water solubility.

Introduction

This kit is suitable for high-throughput extraction of total RNA from paraffin embedded tissue samples, including miRNA. The kit is based on superparamagnetic magnetic particle purification technology. The extraction process does not require the use of toxic phenolic chloroform extraction or time-consuming alcohol precipitation.The whole extraction process only takes 120 minutes. The obtained RNA can be directly used in RT-PCR. The high-throughput extraction can be realized by 96 hole magnetic frame and 96 holeo scillator, various nucleic acid extractors (such as KingFisherFle, MagMix32, MagMix 96) or cooperating with the pipette workstation (Hamilton, Tican) for automatic extraction.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from FFPE tissue
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Products	RNA, miRNA
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Sample type	FFPE slice, FFPE embedded tissue
Sample amount	≤6 slices

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• Fast – several samples can be extracted in 120 minutes by column method
• Safe – no phenol chloroform extraction required

Kit Contents

Contents	IVD3022
Purification Times	200 Preps
MagBind Particles	4.5 ml
Proteinase K	100 mg
Protease Dissolve Buffer	6 ml
DNase I	4 x 600 µl
DNase Buffer	30 ml
Buffer DPS	200 ml
Buffer FRL	40 ml
Buffer AL	40 ml
Buffer MW1*	110 ml
Buffer MW2*	2 x 50 ml
Nuclease Free Water	30 ml

Storage and Stability

Proteinase K and MagBind Particles should be stored at 2–8°C upon arrival. DNase I should bestored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K and MagBind Particles up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under theseconditions.

This kit is suitable for high-throughput extraction of total RNA from paraffin embedded tissue samples, including miRNA. The kit is based on superparamagnetic magnetic particle purification technology. The extraction process does not require the use of toxic phenolic chloroform extraction or time-consuming alcohol precipitation.The whole extraction process only takes 120 minutes. The obtained RNA can be directly used in RT-PCR. The high-throughput extraction can be realized by 96 hole magnetic frame and 96 holeo scillator, various nucleic acid extractors (such as KingFisherFle, MagMix32, MagMix 96) or cooperating with the pipette workstation (Hamilton, Tican) for automatic extraction.