This product is not tested specifically for use in cell culture, however, the product information provided for C9697 may be applied to this product. See the link below to review the product data-sheet. For tissue culture applications, collagenase can be solubilized in calcium-free solutions such as Hank's Balanced Salts (Cat. No. H 2387) or Earles Balanced Salt Solution (Cat. No. E6267).
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/390/021/c9697pis-ms.pdf
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應用
本产品已用于胰腺导管腺癌细胞系的分离和胰腺成纤维细胞的组织培养。[1]
梭菌肽酶A在一项研究中被用来描述一种基于MALDI-TOF（基质辅助激光解吸/电离飞行时间）质谱（MS）法评估胶原蛋白消化后少量胶原蛋白的定量方法。
生化／生理作用
从组织中进行细胞的有效释放需要胶原酶和中性蛋白酶的作用。胶原酶被每克摩尔酶 4 克原子钙（Ca2+）活化。培养滤液被认为含有至少7种不同蛋白酶，分子量范围为68-130 kDa。最适pH为6.3-6.8。该酶通常用于消化组织样品中的结缔组分以释放单个细胞。胶原酶治疗可导致一些细胞死亡。通常，使用0.1至5 mg/mL的浓度用于消化。为产生令人满意的细胞解离效率并不会造成太多的细胞死亡，其反应时间从15分钟到几个小时不等。最好使用含有钙和BSA的Krebs Ringer缓冲液，Zn2+ 是活性必需的。
每摩尔胶原蛋白酶被4g原子钙活化。 它被乙二醇-双（β-氨基乙基醚）-N，NN′,N′-四乙酸，β-巯基乙醇、谷胱甘肽、巯基乙酸和8-羟基喹啉抑制。
品質
也具有梭菌蛋白酶、非特异性中性蛋白酶和胰蛋白酶活性。
單位定義
一个胶原蛋白消化单元（CDU）从牛跟腱胶原蛋白中释放的肽链量和1.0 μ亮氨酸，5小时，pH7.4,37℃，在钙离子存在下的茚三酮颜色相同。一个 FALGPA水解单元每分钟，25℃，水解1.0 μmole 呋喃基丙烯酰-Leu-Gly-Pro-Ala。一个中性蛋白酶单元水解酪蛋白，产生颜色和1.0 μmole 酪氨酸每5小时，pH7.5,37℃下产生的相同。一个梭菌蛋白酶单元在DTT存在下，pH7.6,25℃，每分钟水解1.0 μmole BAEE。
準備報告
可通过向含有0.36mM氯化钙（TESCA），pH为7.4，37℃的50mM TES缓冲液中溶解0.05-0.1mg/mL的胶原酶而制备贮存液。
基底
訊號詞
Danger
危險分類
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
標靶器官
Respiratory system
儲存類別代碼
11 - Combustible Solids
水污染物質分類（WGK）
WGK 1
閃點（°F）
Not applicable
閃點（°C）
Not applicable
個人防護裝備
dust mask type N95 (US), Eyeshields, Faceshields, Gloves
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分析證明 (COA)
Lot/Batch Number
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條款
To measure collagenase activity, N-(3-[2-Furyl]acryloyl)-Leu-Gly-Pro-Ala is used in a continuous spectrophotometric rate determination at 345 nm. Collagenase hydrolyzes collagen peptide bonds.
為了測量膠原蛋白酶的活性，使用 N-(3-[2-糠基]丙烯酰基)-Leu-Gly-Pro-Ala 在 345 nm 下進行連續分光光度速率測定。膠原蛋白酶水解膠原蛋白肽鍵。
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How can I dissolve this product for Tissue digestion (Hanks Buffer or even medium possible)? Thanks
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