柠檬酸（CA）检测试剂盒
产品名称： 柠檬酸（CA）检测试剂盒
英文名称： CITRIC ACID (CA) detection kit
产品编号： IS106
产品价格： 1
产品产地： null
品牌商标： null
更新时间： null
使用范围： null
武汉云克隆诊断试剂研究所
- 联系人 :
- 地址 : 武汉经济技术开发区出口加工区
- 邮编 : 430056
- 所在区域 : 湖北
- 电话 : 138****2189 点击查看
- 传真 : 点击查看
- 邮箱 : 304061971@qq.com
[ INTENDED USE ]
The kit is a colorimetric method for the in vitro quantitative measurement of Citric Acid in samples.
Citric acid (CA), an organic acid commonly found in living organisms, is an important food flavoring substance. In addition, CA is the product of the first reaction of the tricarboxylic acid cycle.
[ REAGENTS AND MATERIALS PROVIDED ]
[ MATERIALS REQUIRED BUT NOT SUPPLIED ]
1. A visible spectrophotometer capable of measuring absorbance at 545nm
2. Thermostatic water bath capable of controlling temperature at 30℃
3. High-speed freezing centrifuge
4. Micropipets and tips
5. 1 mL glass cuvette
6. Distilled water (preferably double distilled water and double distilled water)
[ STORAGE OF THE KITS ]
Reagent 1: liquid. Can be stored at 4℃ for 6 months.
Reagent 2: liquid. Can be stored at 4℃ for 6 months.
Reagent 3: liquid. Can be stored at -20℃ for 6 months.
Reagent 4: powder. Can be stored at room temperature for 6 months.
Reagent 5: liquid. Can be stored at -20℃ for 6 months. Away from light.
Standard: 250μmol/L Citric acid standard solution, liquid. Can be stored at 4℃ for 6 months.
[ REAGENT PREPARATION ]
Reagent 4 Preparation: Before use, add 5ml reagent 1 to dissolve the powder completely.
[ SAMPLE PREPARATION ]
1. Citric acid extraction from liquid sample
Take 0.1 ml of liquid and add 0.9ml of reagent 1, mix well. 11000 g, centrifuge the sample at 10℃, 11000g for 10 min. Extract the supernatant on ice for further measurement.
2. Citric acid extraction from tissue sample
Weight the sample precisely and add the reagent 1 with ratio of 1g sample with 4-9ml of reagent 1(w:v = 1:5-1:10, e.g. 1mL of reagent 1 is added in 0.1g tissue sample). Homogenize the mixture in the ice water bath and the centrifuge the homogenate at 4℃, 11000 g for 10min. Extract the supernatant on ice for further measurement.
3. Citric acid extraction from mitochondria sample
Weight the sample precisely and add the reagent 1 with ratio of 1g sample with 4-9ml of reagent 1(w:v = 1:5-1:10, e.g. 1mL of reagent 1 is added in 0.1g tissue sample). Homogenize the mixture in the ice water bath and the centrifuge the homogenate at 4℃, 600 g for 5min. Collect the supernates in another EP tube and centrifuge it at 4℃, 11000 g for 10min. Remove the supernatant and then add 200ul of reagent 2 and 2ul of reagent 3. after on ice for further measurement. Mix sufficiently and centrifuge it at 4℃, 11000 g for 10min. Extract the supernatant on ice for further measurement.
4. Citric acid extraction from bacteria and fungi sample.
Add the reagent 1 with ratio of 500-1000×104 cells with 1ml of reagent 1(e.g. 1mL of reagent 1 is added in 500×104 cells). The cells could be subjected to ultrasonication till the solution is clarified(e.g. Ultrasonic power: 300W, ultrasonic:3 seconds, Stop:7 seconds, total time: 3 minutes). Centrifuge at 4℃, 11000g for 10 minutes to remove cellular debris. Extract the supernatant on ice for further measurement.
[ ASSAY PROCEDURE ]
Operation table:
Blank tube
Standard tube
Sample tube
Distilled Water（μL）
Note: Before use, the reagent should be preheated in a 30℃ water bath for 30 min.
[ TEST PRINCIPLE ]
Under acidic conditions, citric acid reduced Cr6+ to form Cr3+, and Cr3+ had a characteristic absorption peak at 545nm. By measuring the increase of 545nm absorbance, the citric acid content in the sample can be calculated.
[ CALCULATION OF RESULTS ]
[ IMPORTANT NOTE ]
1. Samples pre-treatment should be carried out on ice.
2. Prepare reagent 4 within 15 minutes before assay. The reconstituted reagent 4 can be used only once.
3. Reagent 5 contains carcinogens. During the experiment, wear gloves to prevent skin exposure.
4. The citric acid extract buffer can not be used for the determination of protein concentration. If needed, it is recommended to take an additional tissue and use IS003 BCA Protein Quantification Kit to determine the protein concentration.
The kit is a colorimetric method for the in vitro quantitative measurement of Citric Acid in samples.
Citric acid (CA), an organic acid commonly found in living organisms, is an important food flavoring substance. In addition, CA is the product of the first reaction of the tricarboxylic acid cycle.
[ REAGENTS AND MATERIALS PROVIDED ]
[ MATERIALS REQUIRED BUT NOT SUPPLIED ]
1. A visible spectrophotometer capable of measuring absorbance at 545nm
2. Thermostatic water bath capable of controlling temperature at 30℃
3. High-speed freezing centrifuge
4. Micropipets and tips
5. 1 mL glass cuvette
6. Distilled water (preferably double distilled water and double distilled water)
[ STORAGE OF THE KITS ]
Reagent 1: liquid. Can be stored at 4℃ for 6 months.
Reagent 2: liquid. Can be stored at 4℃ for 6 months.
Reagent 3: liquid. Can be stored at -20℃ for 6 months.
Reagent 4: powder. Can be stored at room temperature for 6 months.
Reagent 5: liquid. Can be stored at -20℃ for 6 months. Away from light.
Standard: 250μmol/L Citric acid standard solution, liquid. Can be stored at 4℃ for 6 months.
[ REAGENT PREPARATION ]
Reagent 4 Preparation: Before use, add 5ml reagent 1 to dissolve the powder completely.
[ SAMPLE PREPARATION ]
1. Citric acid extraction from liquid sample
Take 0.1 ml of liquid and add 0.9ml of reagent 1, mix well. 11000 g, centrifuge the sample at 10℃, 11000g for 10 min. Extract the supernatant on ice for further measurement.
2. Citric acid extraction from tissue sample
Weight the sample precisely and add the reagent 1 with ratio of 1g sample with 4-9ml of reagent 1(w:v = 1:5-1:10, e.g. 1mL of reagent 1 is added in 0.1g tissue sample). Homogenize the mixture in the ice water bath and the centrifuge the homogenate at 4℃, 11000 g for 10min. Extract the supernatant on ice for further measurement.
3. Citric acid extraction from mitochondria sample
Weight the sample precisely and add the reagent 1 with ratio of 1g sample with 4-9ml of reagent 1(w:v = 1:5-1:10, e.g. 1mL of reagent 1 is added in 0.1g tissue sample). Homogenize the mixture in the ice water bath and the centrifuge the homogenate at 4℃, 600 g for 5min. Collect the supernates in another EP tube and centrifuge it at 4℃, 11000 g for 10min. Remove the supernatant and then add 200ul of reagent 2 and 2ul of reagent 3. after on ice for further measurement. Mix sufficiently and centrifuge it at 4℃, 11000 g for 10min. Extract the supernatant on ice for further measurement.
4. Citric acid extraction from bacteria and fungi sample.
Add the reagent 1 with ratio of 500-1000×104 cells with 1ml of reagent 1(e.g. 1mL of reagent 1 is added in 500×104 cells). The cells could be subjected to ultrasonication till the solution is clarified(e.g. Ultrasonic power: 300W, ultrasonic:3 seconds, Stop:7 seconds, total time: 3 minutes). Centrifuge at 4℃, 11000g for 10 minutes to remove cellular debris. Extract the supernatant on ice for further measurement.
[ ASSAY PROCEDURE ]
Operation table:
Blank tube
Standard tube
Sample tube
Distilled Water（μL）
Note: Before use, the reagent should be preheated in a 30℃ water bath for 30 min.
[ TEST PRINCIPLE ]
Under acidic conditions, citric acid reduced Cr6+ to form Cr3+, and Cr3+ had a characteristic absorption peak at 545nm. By measuring the increase of 545nm absorbance, the citric acid content in the sample can be calculated.
[ CALCULATION OF RESULTS ]
[ IMPORTANT NOTE ]
1. Samples pre-treatment should be carried out on ice.
2. Prepare reagent 4 within 15 minutes before assay. The reconstituted reagent 4 can be used only once.
3. Reagent 5 contains carcinogens. During the experiment, wear gloves to prevent skin exposure.
4. The citric acid extract buffer can not be used for the determination of protein concentration. If needed, it is recommended to take an additional tissue and use IS003 BCA Protein Quantification Kit to determine the protein concentration.