Solarbio ABTS Tree Radical Scavenging Activity Assay Kit

Produktübersicht

Attribut	Einzelheiten
Betriebsausrüstung	Spektrophotometer
Katalognummer	BC4770
Größe	50 Tests / 24 Proben

Product Content:

Please ensure the reagent volume matches the solution volume in the bottle before use. For inquiries, kindly contact Solarbio staff promptly.

Reagenzname	Spezifikationen	Lagerbedingungen
Extract Solution	Flüssig, 60 ml × 1	Speichern Sie bei 4 ℃
Reagenz I	Flüssig, 80 ml × 1	Speichern Sie bei 4 ℃
Reagenz II	Powder × 1	Speichern Sie bei 4 ℃
Reagenz III	Flüssig, 20 μL × 1	Speichern Sie bei 4 ℃
Reagenz IV	Flüssig, 1.5 ml × 1	Bei -20℃ lagern
Reagenz V	Powder × 1	Speichern Sie bei 4 ℃

Preparation of Solution:

1. Reagenz II: Vor Gebrauch, hinzufügen 3 mL of distilled water and fully dissolve. Any unused reagent can be stored at -20℃ for two weeks.
2. Preparation of Reagent III Working Solution: Place the liquid in the provided EP tube. Prepare the working solution according to the ratio of Reagent III (μL) to distilled water (ml) = 1 μL: 12 ml. Unused reagents should be stored at 4℃.
3. Preparation of Reagent IV Working Solution: Store Reagent IV separately at -20℃. Vor Gebrauch, prepare the working solution according to the sample number and the ratio of Reagent V to Reagent I (V: V) = 1:9. Unused reagent should be stored at -20℃.
4. Reagenz V: Place the powder containing 5 mg of vitamin C in the provided EP tube. Hinzufügen 2.8 mL of Extract Solution before use, and shake thoroughly to dissolve. Bereiten a 10 mmol/L vitamin C solution for use as a positive control. Store at 4℃ for one week.
5. Preparation of ABTS Working Solution: Vor Gebrauch, prepare the ABTS working solution according to the required amount for the test. The proportion of Reagent I: Reagenz II: Reagent III Working Solution (V:V:V) = 76:5:4. Prepare and store in a dark room at room temperature. Use within 30 Protokoll.

Produktbeschreibung:

1. Antioxidant Capacity Assessment: The ABTS method is widely used to assess the antioxidant capacity of hydrophilic and lipophilic substances.
2. Formation of ABTS Radical: ABTS oxidizes to form a stable blue-green cation ABTS radical with absorption peaks at 405 nm or 734 nm.
3. Decrease in Absorbance: When the tested substance is added to the ABTS radical solution, the antioxidant component reacts with it, causing a decrease in absorbance at 405 nm.
4. Proportional Scavenging: The change in absorbance is directly proportional to the degree of free radical scavenging within a certain range.
5. Quantification of Scavenging: This kit measures the sample’s ability to scavenge ABTS radicals by quantifying the decrease in absorbance.

Notiz: Before testing, conduct preliminary experiments with 2–3 samples showing significant expected differences. If sample absorbance falls outside the measurement range, consider diluting or increasing the sample size for testing.

Erforderliche, aber nicht bereitgestellte Reagenzien und Ausrüstung:

Constant temperature water bath, spectrophotometer, 1 ML Glass Cuvette, desktop centrifuge, mortar/grinder, drying box, 30~50 mesh sieve, und destilliertes Wasser.

Verfahren:

ICH. Probenextraktion: (The number of samples can be adjusted accordingly, and refer to literature for specific proportions)

1. Preparation of Plant Tissue Samples: Dry fresh samples at 60℃ until constant weight, grind in a mortar (or grinder), and sieve through a 30~50 mesh sieve. Ungefähr wiegen 0.05 g of samples, hinzufügen 1 mL of Extract Solution, and incubate in a 40℃ water bath for 30 Mindest. Zentrifuge bei 10000 U/min für 10 min at room temperature, Sammeln Sie den Überstand und halten Sie ihn zum Testen auf Eis.
2. Red Wine, Fruit Juice, and Other Liquid Samples: Nehmen 100 μL of sample solution and add 900 μL of Extract Solution. Vortex mix, centrifuge at room temperature, 10000 U/min für 10 Mindest, Sammeln Sie den Überstand und halten Sie ihn zum Testen auf Eis.
3. Extraction or Drug: Prepare a certain concentration with the Extract Solution, wie zum Beispiel 5 mg/ml.

Notiz: The ability of various samples to scavenge ABTS free radicals may vary significantly. To ensure accuracy, adjust samples according to preliminary results.

II. Determination Steps:

1. Preheat the spectrophotometer for over 30 Protokoll, set the wavelength to 405 nm, und Null mit destilliertem Wasser.
2. Preparation of Positive Control: Bereiten a 10 mmol/L vitamin C solution with Extract Solution. For a linear relationship, vorbereiten 1, 0.8, 0.6, 0.4, 0.2, Und 0.1 mg/mL vitamin C solutions. For a positive control with a clearance rate of about 90%, prepare vitamin C solutions larger than 1.5 mmol/L.
3. Operational Table: Add the following reagents into a 96-well plate or EP tube, respectively:

Reagenzname	Leeres Rohr (B)	Testrohr (T)	Steuerrohr (C)	Positive Control Tube (PC)
Überstand	50	50		50
VC Solution				50
Destilliertes Wasser	50		50
Reagent IV Solution	100	100	100	100
ABTS Working Solution	850	850	850	850
Reagenz I	950

After thorough mixing, incubate in darkness for 6 min at room temperature. Messen Sie die Absorption bei 405 nm. Record absorbance values for blank tube, control tube, positive control tube, and test tube as AB, AC, APC, and AT respectively. The blank tube needs to be tested 1-2 mal.

III. Calculation of ABTS Free Radical Scavenging Rate:

1. Establishment of Standard Curve: The formula for the free radical scavenging rate of the positive control: ABTS Free Radical Scavenging Rate DVC% = [(AB-APC) ÷ AB] × 100%
2. Calculation Formula of Sample Free Radical Scavenging Rate: ABTS Free Radical Scavenging Rate D% = [AB – (AT-AC)] ÷ AB] × 100%.

Notiz:

1. To compare the scavenging ability of different samples, it is recommended to add the same volume of samples to the same batch, adjusting according to pre-experimental results. During comparison, adjust samples according to results, comparing clearance rates at the same concentration (Verdünnungsverhältnis).
2. Samples should be taken and tested on the same day.

Experimental Example:

Nehmen 0.05 g of chili and add 1 mL of Extract Solution to homogenize. Nach Zentrifugation, collect the supernatant and proceed according to the determination steps. Calculate the clearance rate: D% = [AB – (AT-AC)] ÷AB] × 100% = [1.330-(0.468-0.022)]÷ 1.330×100% = 66.466%

Verwandte Produkte:

• BC4750/BC4755: DPPH Free Radical Scavenging Capacity Assay Kit
• BC1310/BC1315: Total Antioxidant Capacity (T-AOC) Assay-Kit
• BC1320/BC1325: Hydroxyl Radical Scavenging Capacity Assay Kit