膠原蛋白檢測試劑盒 | EnzyFluo^TM Collagen Assay Kit  BioAssay Systems 貨號ECOL-100

膠原蛋白(COLLAGEN)是結締組織(connective tissue)的關鍵結構蛋白(structural protein)，也是哺乳動物(mammals)體內最豐富的蛋白質。它以許多不同的類型和形式出現，其中 I-V 型(Types I -V)最為常見。除了它在身體中發揮的關鍵作用外，它還有許多醫學應用，例如用於骨(bone)和皮膚移植(skin grafts)等重建手術(reconstructive surgery)。由於其抗衰老(anti-aging)和皮膚癒合(skin healing)特性，它也常用於化妝品中。目前可用於量化膠原蛋白的測定方法範圍從需要用酸和鹼進行廣泛的水解程序到使用昂貴的抗體和複雜的方案。 BioAssay Systems 的膠原蛋白檢測試劑盒(BioAssay Systems’ collagen assay kit)提供了一種非常簡單、安全且靈敏的方法來量化樣品中的膠原蛋白(collagen)。在此過程的第一步中，樣品中的膠原蛋白被酶(enzyme)消化成肽(peptides)。隨後，含有肽(peptides)的 N 端甘氨酸(N-terminal glycine)與染料反應形成螢光複合物(fluorescent complex)。該產品的螢光強度（在 λex/em = 375/465 nm 處測量）與樣品中的膠原蛋白(collagen)濃度成正比。

 

Application

• For quantitatve determination of collagen in biological and cosmetic products.

Key Features

• Non-radioactive assay.
• Fast and sensitive. Linear detection range (20µL sample): 2 µg/mL to 50µg/mLcollagen in 96-well plate assay.
• Convenient and high-throughput.”Add-mix-read” type assay. No wash and reagent transfer steps are involved. Can be readily automated for processing thousands of samples per day.

Method

• FL375/465nm

Samples

• Biological samples (e.g. plasma, serum, cells) and cosmetic samples (e.g. collagen solution)

Species

• All

Procedure

• 80 min

Size

• 100 tests

Detection Limit

• 2 µg/mL

Shelf Life

• 6 months

BioAssay Systems 品牌介紹

BioAssay Systems 公司研製開發和銷售創新(innovative)高通量的檢測試劑組(high-throughput assay solutions)，以滿足不斷增加的需求的生命科學行業(life sciences industry)。BioAssay Systems 專注於安全，無放射性等格式的吸收光，螢光和發光檢測技術。BioAssay Systems 在現代藥物發現方面擁有 50 多年的豐富經驗。在過去的 18 年中，開發 200 多種用於藥物發現和生命科學研究的檢測試劑盒。提供的服務和簡單、使用方便且性能卓越的產品而自豪。

BioAssay Systems檢測試劑盒及試劑

• Agriculture & Environment
• Blood & Urine Chemistry
• Cations & Anions
• Diabetes & Obesity
• Enzyme Activity
• Food & Beverage Analysis
• HTS Reagents & Kits
• Inhibitor HTS Kits
• Metabolism
• Oncology
• Other Reagents
• Oxidative Stress
• Quick Test Strips
• Signal Transduction

Does this assay measure hydroxyproline?

No, this assay does not measure hydroxyproline. This assay quantifies N-terminal glycine containing peptides from an enzyme based degradation of collagen. We have a hydroxyproline assay, DHYP-100.

My standard curve isn’t turning out linear / I am getting inconsistent results?

You may also wish to incubate the digest mix step for an extra 30 minutes at 37°C (90 minutes total) to ensure that the digest reaction is complete.

I’m not getting the same fluorescence readings for my standards as the standard curve on the protocol?

Fluorescence is measured differently depending on the plate reader. This is why plate readers use relative fluorescent units (RFUs) to quantitate the fluorescence readings. These are arbitrary units to assess the fluorescence intensity of the wells relative to each other. The numbers you see on the standard curve in the protocol are from the assay being performed on our lab’s plate readers. It is completely normal for your assay to produce different fluorescent readings; what is more important is to focus on the linearity of your standard curve.

My sample, which I know has collagen, isn’t measuring any collagen with this assay?

Either your samples are diluted too much, or have not been diluted enough. We recommend running a serial dilution on your sample to determine the optimal dilution factor for quantification. This is the range at which dilutions start producing similar collagen concentration estimates after being corrected for dilution factor.

Can I use this assay to measure cosmetic or beauty product collagen?

Yes, this assay can be used on store bought collagen. Because the collagen is often very concentrated, you will likely need to dilute it greatly (e.g.100-1000 fold) for it to be within the assay’s detection range.

Hong, S., et al. (2020). Ginsenoside compound-Mc1 attenuates oxidative stress and apoptosis in cardiomyocytes through an AMP-activated protein kinase-dependent mechanism.Journal of Ginseng Research.44(4): 664-671. Assay: Collagen in mouse heart tissue.