酪氨酸酶活性測定試劑盒 | Tyrosinase Activity Assay Kit ATTOGENE貨號EZ2017-01 (30 µU)

For rapid, sensitive and accurate screening of potential Tyrosinase inhibitors

酪氨酸酶 (Tyrosinase, EC 1.14.18.1) 是一種銅結合酶(copper-binding enzyme )，在從細菌(bacteria )和真菌(fungi )到哺乳動物( mammals)的廣泛物種中都有表達。它參與黑色素合成途徑(melanin synthesis pathway)的兩個連續反應；首先是單酚(monophenol )的羥基化(hydroxylation)，其次是鄰二酚向醌(ortho-diphenol )的轉化。醌然後經歷一系列反應，包括聚合(polymerization)形成黑色素(melanin)。黑色素（melanin）在黑色素細胞（melanocytes）的生物合成的途徑是由酪胺酸 （tyrosine）與酪胺酸酶（tyrosinase）的作用開始。現今已知的美白成分之中，對於酪胺酸酶活性抑制的專一抑制劑，通常具有與酪胺酸相似的結構，在黑色素細胞內進行 競爭性抑制酪胺酸酶以達減少黑色素生成。

酪氨酸酶(Tyrosinase )引起農業工業的極大興趣，因為它會引起水果(fruits)、蔬菜(vegetable )和蘑菇(mushrooms)的褐變，以及引起皮膚變黑(skin darkening)的化妝品工業。因此，酪氨酸酶抑製劑( tyrosinase inhibitors)的開發和篩選(screening)對於色素沉著過度(hyperpigmentation)和黃褐斑(melasma)等病症非常有用。黑色素瘤(melanoma)中的酪氨酸酶活性(Tyrosinase activity )顯著增加。因此，檢測酪氨酸酶活性(tyrosinase activity )有望成為黑色素瘤(melanoma)的特異性診斷測試(specific diagnostic test )，並可用於監測患者對黑色素瘤治療(melanoma treatments)的反應。 environmental, Food Safety, life science, pesticide, Tyrosinase Activity

酪氨酸酶活性測定原理介紹

酪氨酸酶活性測定試劑盒(Tyrosinase Activity Assay)是一種簡單的基於板的一步式測定法，用於測量生物樣品中的酪氨酸酶活性( tyrosinase activity)。在此測定中，酪氨酸酶(tyrosinase)催化酚類底物轉化為醌中間體(Quinone intermediate)，後者與酪氨酸增強劑(tyrosine enhancer)反應，形成高度穩定的髮色團(chromophore)，在 520 nm 處有吸光度。該測定法可檢測生物樣品中低至 30 µU 的酪氨酸酶( Tyrosinase )。

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 inter-mediate, 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.

The kit provides a rapid, simple, sensitive, and reliable test suitable for screening of tyrosinase concen-tration in samples.

該試劑盒提供了一種快速(rapid)、簡單(simple)、靈敏(sensitive)且可靠的檢測方法，適用於樣品中酪氨酸酶濃度(tyrosinase concen-tration)的 screening。酪氨酸酶濃度可以通過讀取反應在 520nm 處的吸光度來測量，使用試劑盒中提供的標準( standards)生成標準曲線(standard curve)並使用線性回歸分析未知樣品的濃度。

Set up reaction in 96 well plate from the master mix by:

• Step 1. Aliquoting 223μL of the master mix into each well of the 96 well plate.
• Step 2. Add 40μL of Sample, Tyrosinase Lysis Buffer or Tyrosinase Positive Control
• Step 3. Add 7μL of Substrate [S]
• Step 4. Mix the components in the tube by pipetting up and down 3-4 times
• Step 5. Incubate at room temperature for 25 minutes.
• Step 6. Read the plate by measuring absorbance at 520nm.

Concentration Calculation:
Standard curve can be constructed by plotting the mean relative absorbance (%) obtained from each reference standard against its concentration in units on a logarithmic curve.
Relative absorbance (%) = absorbance standard (or sample) x 100 absorbance zero standard
Use the mean relative absorbance values for each sample to determine the corresponding concen-tration of the sample from the standard curve.

Cat # +Size	EZ2017-01
Size	100 assays
Kit Summary	• Detection method- Absorbance (520 nm) • Applications- Measurement of Tyrosinase Activity in various tissue/cell lysates – Analysis of Tyrosinase activity in food chemistry/skin-related research
Detection Method	Absorbance (520 nm)
Applications	• Measurement of Tyrosinase Activity in various tissue/cell lysates • Analysis of Tyrosinase activity in food chemistry/skin-related research
Features & Benefits	• First non-radioactive assay in the market • Simple, rapid & convenient • The assay can detect Tyrosinase Activity less than 30 µU in biological samples.

Attogene 台灣代理/Attogene 台灣經銷

10nm Colloidal Gold、30nm Colloidal Gold、 40nm Colloidal Gold 、

Attogene 提供側流分析開發(Lateral Flow Assay)奈米金粒子( Colloidal Gold)

用於橫向流動(Lateral Flow Assay)的膠體金( Colloidal Gold)是一種高度穩定且均勻的金納米粒子(gold nanoparticles)，膠體金( Colloidal Gold)用於抗體的傳統被動結合， 無塗層金納米粒子於Passive conjugation被動共軛進行了優化，可應用於 lateral flow tests側向流動測試。提供形狀尺寸分佈最平均之奈米金粒子。適用於：蛋白質吸附、寡核苷酸吸附、表面修飾。Attogene奈米金溶液特點: 形狀最一致、尺寸分佈最平均10nm ± 1.5nm，均勻的形狀確保抗體的一致結合，確保可靠的分析結果。高抗體結合率以達最佳靈敏度。每批10nm膠體金: 平均直徑：8.5-11.5nm

可提供 1 OD 至 100 OD 的範圍(粒子濃度2.5- 8.6 x 10¹²/mL)。偶聯物( conjugate )的質量和性能對於成功的橫向流動測試生產(lateral flow test manufacturing)至關重要。ATTOGENE產品在美國製造，並在最先進的製造設施中生產，可實現快速周轉時間，同時確保批次間的一致性和可靠性。建議的儲存和處理程序：儲存在 2 – 25^oC. 不要凍結。冷凍會導致不可逆的聚集顆粒並破壞產品。置於室溫並劇烈搖晃 30 秒以確保顆粒充分每次使用前分散使用。每ml的奈米金粒子2.5- 8.6 x 10¹²/mL。膠體金濃度 (mg/mL)為5.7- 6.0 x 10^-2。Molar Concentration (moles/liter)0.4-1.4 x 10^-8

Number of particles/mL	5.2-7.2 x 1010
Gold Concentration (mg/mL)	3.7-4.2 x 10-2
Molar Concentration (moles/liter)	8-12 x 10-11
Particle Diameter	40 nm /- 1.5

Related products:

AU2012-01: 10nm Colloidal Gold (1O.D, 1L) >>操作手冊

AU2013-01: 30nm Colloidal Gold (1O.D, 1L) >>操作手冊

AU2014-01: 40nm Colloidal Gold (1O.D, 1L) >>操作手冊

AU2015-01: 60nm Colloidal Gold (1O.D, 1L) >>操作手冊

40nm奈米金溶液特點: (1O.D/1公升, 貨號AU2014-01)

形狀最一致、尺寸分佈最平均40nm ± 2.5nm，均勻的形狀確保抗體的一致結合，確保可靠的分析結果。高抗體結合率以達最佳靈敏度。每批40nm膠體金(1 O.D): 平均直徑：37.5-42.5nm

40 nm奈米金顆粒(1O.D/1公升, 貨號AU2014-01)的紫外-可見光譜

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10nm奈米金溶液特點: (1O.D/1公升, AU2012-01)

形狀最一致、尺寸分佈最平均10nm ± 1.5nm，均勻的形狀確保抗體的一致結合，確保可靠的分析結果。高抗體結合率以達最佳靈敏度。每批10nm膠體金(1 O.D): 平均直徑：8.5-11.5nm、

10 nm奈米金顆粒  (1O.D/1公升, AU2012-01)的紫外-可見光譜

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30nm奈米金溶液特點: (1O.D/1公升, 貨號AU2013-01)

形狀最一致、尺寸分佈最平均30nm ± 1.5nm，均勻的形狀確保抗體的一致結合，確保可靠的分析結果。高抗體結合率以達最佳靈敏度。每批30nm膠體金(1 O.D): 平均直徑：28.5-31.5nm

Specifications 1OD 30nm Gold (1L)

• Number of particles/mL	1.3-2 x 1011
• Gold Concentration (mg/mL)	4.2-4.7 x 10-2
• Molar Concentration (moles/liter)	2.2-3.3 x 10-10
• Particle Diameter	30 nm /-1.5

30 nm奈米金顆粒  (1O.D/1公升, AU2013-01)的紫外-可見光譜

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60nm奈米金溶液特點: (1O.D/1公升, 貨號AU2015-01)

形狀最一致、尺寸分佈最平均60nm ± 4.5nm，均勻的形狀確保抗體的一致結合，確保可靠的分析結果。高抗體結合率以達最佳靈敏度。每批60nm膠體金(1 O.D): 平均直徑：65.5-64.5nm

Specifications 1OD 60nm Gold (1L)

Number of particles/mL	1.5-2 x 1010
Gold Concentration (mg/mL)	3.6-4.1 x 10-2
Molar Concentration (moles/liter)	2.5-3.4 x 10-11
Particle Diameter	40 nm /- 1.5

60 nm奈米金顆粒  (1O.D/1公升, AU2015-01)的紫外-可見光譜

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ATTOGENE提供 1 OD 至 100 OD 的範圍(以40nm奈米金粒子為例，粒子濃度2.5- 8.6 x 10¹²/mL)。偶聯物( conjugate )的質量和性能對於成功的橫向流動測試生產(lateral flow test manufacturing)至關重要。

建議的儲存和處理程序：

儲存在 2 – 25^oC. 不要凍結。冷凍會導致不可逆的聚集顆粒並破壞產品。置於室溫並劇烈搖晃 30 秒以確保顆粒充分每次使用前分散使用。以40nm奈米金粒子為例，每ml的奈米金粒子2.5- 8.6 x 10¹²/mL。膠體金濃度 (mg/mL)為5.7- 6.0 x 10^-2。Molar Concentration (moles/liter)0.4-1.4 x 10^-8

奈米金（又稱為膠體金），常以溶液型態呈現。當奈米金粒子越小（<100nm）則呈現紅色溶液。反之（>100nm）則越偏向藍紫色。奈米金粒子用途廣泛，可應用於橫向流分析、顯微影像、流式細胞、萊曼光譜、以及動態光散射（DLS）。在這些應用中，奈米金粒子的大小是非常重要的影響關鍵。雖然奈米金的來源多種（自製or商品化產品），ATTOGENE產品在美國製造，並在最先進的製造設施中生產，可實現快速周轉時間，同時確保批次間的一致性和可靠性。

Colloidal gold applications

Application	Size				Shape
	10	20	40	80
Lateral Flow		●	●	●	Sphere
Western Blots		●	●	●	Sphere
Dot Blots			●		Sphere
Dark Field microscopy			●		Sphere
DLS	●	●	●	●	Sphere
Flow Cytometry			●	●	Sphere
Electron microscopy	●	●			Sphere
Metal enhanced fluorescence			●		Sphere/flat surface
SERS			●	●	Not spherical
FRET quench	●	●	●		Sphere
Infrared phototermal therapy		●	●		Rod/hollow sphere
Radiofrequency ablation			●		Sphere
Drug delivery	●	●	●		Rod/sphere