Collagenase, Type I, powder

Name: Collagenase, Type I, powder
SKU: 17100017

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製品番号（カタログ番号）	数量
17100017	1 g
17018029	500 mg

2 オプション

製品番号（カタログ番号） 17100017

価格（JPY）

54,500

Each

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数量:

1 g

コラゲナーゼは、中性アミノ酸（X）とグリシンの間の結合を分解するプロテアーゼで、コラーゲン内で高い頻度で検出されるPro-X-Gly-Pro配列に含まれます。コラゲナーゼは、皮膚、腱腱、血管、骨などの結合組織で一般的に見られる三重らせん天然コラーゲン原線維を分解できる点でプロテアーゼの中で特異的です。コラゲナーゼ分解は、ヒト腫瘍、マウス腎臓、ヒトの成人および胎児の脳、および上皮を含む多くの他の組織の培養に適しています。コラゲナーゼは比較的穏やかで、生理学的温度とpHで良好に解離し、機械的撹拌や特別な装置は必要ありません。

Gibco™ コラゲナーゼI型はヒストリチクム菌から単離され、クロストリジウム組織ライティクムから単離され、細胞または組織の解離、および器官の灌流における研究用の凍結乾燥した未滅菌粉末として包装されています。Gibco™ コラゲナーゼI型活性は、125ユニット／mgを超えることが保証されています。他のコラゲナーゼ調製と比較して、Gibco™コラゲナーゼI型は平均濃度のコラゲナーゼ、カセイナーゼ、クロストリパイン、およびトリプシン活性を含み、脂肪、副腎、肝細胞または組織の消化に最適です。

研究用にのみ使用できます。診断用には使用いただけません。

仕様

数量1 g

品質保持期間24 Months

出荷条件Room Temperature

形状Lyophilized

製品タイプCollagenase

無菌性Non-sterile

Unit SizeEach

組成および保存条件

Storage conditions: 2°C to 8°C. Protect from light.
Shipping conditions: Room temperature
Shelf life: 24 months from date of manufacture

よくあるご質問（FAQ）

Can collagenase be used to dissociate feeder free hESC when cultured on CELLstart medium?

Yes, collagenase can be used. It is recommended to use a 10 mg/ml collagenase IV solution (0.5 ml per 6 cm dish). After 1 minute, remove collagenase and rinse cells. Scrape cells into wash buffer (see product manual for more information), pellet, and re-plate as desired.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

How do I make the 1000X stock (100 U/µL) solution from Collagenase powder?

1. Add 1 mL Hanks' Balanced Salt Solution (HBSS) with calcium and magnesium directly to 1 g vial of Collagenase. Vortex gently to ensure complete dissolution. Transfer to a clean tube.
2. Determine volume of HBSS (with calcium and magnesium) required to bring collagenase solution to 100 U/µL (1000X stock solution). The activity is lot- specific. Rinse vial with this volume of HBSS (with calcium and magnesium), and combine. Filter sterilize 1000X stock solution with a low protein binding filtration unit.
Example: Assuming the lot you have purchased has an activity of 265 U/mg, this lot will have 265000 Units per mL when you reconstitute collagenase into HBSS (with calcium and magnesium) at 1 g/mL. In order to dilute 265000 U/L to 100000 U/mL (= 100 U/µL), you need to dilute the 1 g/mL enzyme solution 2.65 fold.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

What reagents do you offer for cell dissociation, and what are the main differences between them?

Please use this selection chart that compares our cell dissociation reagents (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/reagents/trypsin.html).

Find additional tips, troubleshooting help, and resources within ourMammalian Cell Culture Basics Support Center.

引用および参考文献 (1)

引用および参考文献

Abstract

FRET measurements of intracellular cAMP concentrations and cAMP analog permeability in intact cells.

Authors:Börner S, Schwede F, Schlipp A, Berisha F, Calebiro D, Lohse MJ, Nikolaev VO,

Journal:Nat Protoc

PubMed ID:21412271

'Real-time measurements of second messengers in living cells, such as cAMP, are usually performed by ratiometric fluorescence resonance energy transfer (FRET) imaging. However, correct calibration of FRET ratios, accurate calculations of absolute cAMP levels and actual permeabilities of different cAMP analogs have been challenging. Here we present a protocol that ... More