病人尿液及血液中Creatine含量分析，BIOVISION Creatine Assay Kit，#k635-100; Creatinine assay kit,#K625-100

Creatine Assay Kit
Catalog#: K635-100 | Size: 100 assays

【供應廠商】萊恩生物科技有限公司

【公司電話】04-2523-9639

【業務專員】陳俊豪 0937-229803

【電子信箱】biolion0819@gmail.com

Catalog#	Product Name	Size
K635-100	Creatine Assay Kit	100 assays
K625-100	Creatinine Assay Kit	100 assays
K636-100	Sarcosine Assay Kit	100 assays

Kit Summary:
• Detection method- Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
• Sample type- Cell and Tissue culture supernatants, urine, plasma and serum, as well as many other biological fluids
• Species reactivity- Mammalian
• Applications-The kit can detect 0.001 – 10 mM Creatine.
Features & Benefits:
• Simple procedure; takes ~ 1 hour
• Fast and convenient
• Kit contains all necessary reagents for accurate measurement of Creatine
Kit components:
• Creatine Assay Buffer
• Creatine Probe
• DMSO (Anhydrous)
• Creatinase
• Creatine Enzyme mix
• Creatine Standard (10 µmol)
Description:
Creatine is an endogenous compound whose function is to maintain a high ATP/ADP ratio, by way of its phosphorylated form and creatine kinase. Creatine supplementation has been used in the treatment of muscular, neurological and neurodegenerative diseases, as well as a sport performance enhancer. Detection of creatine level has wide applications in research and development. BioVision’s Creatine Assay Kit provides an accurate, convenient measure of creatine in a variety of biological samples. In the assay, creatine is enzymatically converted to sarcosine which is then specifically oxidized to generate a product that converts a colorless probe to an intensely red color (λmax = 570 nm), and highly fluorescent (Ex/Em = 538/587 nm) product. Creatine is therefore easily detected by either colorimetric or fluorometric methods. Detection range 0.001 – 10 mM Creatine.

1. 尿液及血液中Creatine分析

(1). 方法參照BIOVISION套組 (貨號#K635-100, 太鼎生技)。以570nm波段測量，得到O.D值 (如表2所示)。

(2). 每個O.D.值，需扣除background值 (如表3所示)。(此backgroung control為參照BIOVISION套組內建議)

*Note: Sarcosine gives background for the assay. For samples which may contain a significant amount of sarcosine, do a background control. Prepare a reaction without the creatinase (replacing the creatinase with 2 μl assay buffer).

(3). 以迥歸曲線換算，得到Creatine含量，其單位以nmol表示，其線性範圍介於0~10nmol (如表4所示)。

(4). 換算出Creatine (nmol) 含量後，此Creatine (nmol)含量是較高的，因為尿液及血液樣品，全部經由10Kd spin column濃縮過，所以最後到的Creatine (nmol)讀值會比較高，正確讀值需再除以濃縮倍數，才得到正確的Creatine含量nmol。

因為Creatine含量再除以濃縮倍數後，其讀值會太小，線性範圍 (0~10nmol)，所以最後以最終濃度uM或mM來表示(如表5所示)，最終濃度換算公式如下。

樣品Creatine含量 (nmol) / 50ul = mM

樣品Creatine含量 (nmol) / 50ul *1000 = uM

因為標準品的最終濃度為0~200uM (理論上的值), 所以採用uM為單位

實驗上測出算出標準品的最終濃度為-8.63306~ 189.884uM。