Malondialdehyde Assay Kit (Microanalysis)

• Rich in surface-active functional groups: Magrose features a high density of COOH groups (~1000 μmol/g), ensuring efficient target binding and extremely low nonspecific adsorption.

• Uniform magnetic bead dispersion with good operability, maintaining stable magnetic responsiveness and excellent resuspension performance.

• Excellent physicochemical stability and good batch-to-batch reproducibility; the coefficient of variation (CV) of amino group content between batches is <5%, ensuring stable and reliable experimental results.

• High target binding capacity with low nonspecific adsorption, specifically designed for separation and purification applications.

Taking 100T/96S packing for example:

Before the formal assay, perform a pilot test using 2–3 samples with expected large differences.

1. Preparation of Lab Instruments

Visible spectrophotometer / microplate reader, micro glass cuvettes / 96-well plates, water bath, benchtop centrifuge, adjustable micropipettes, mortar, ice, and distilled water.

2. MDA Extraction Solution

1)Preparation of bacterial and cell samples:

Bacteria or cultured cells: Collect the bacteria or cells into centrifuge tubes. After centrifugation, discard the supernatant. Add CB0094M-ES according to the ratio of bacterial or cell number (10⁴ cells): CB0094M-ES volume (mL) = 500~1000 : 1 (recommended: add 1 mL of CB0094M-ES to 5 x 10⁶ bacteria or cells). Lyse the bacteria or cells by ultrasonication (on ice; 20% power or 200 W; sonicate for 3 s with 10 s intervals, repeat 30 times). Centrifuge at 8000 × g for 10 min at 4 °C. Collect the supernatant and keep it on ice for analysis.

2)Preparation of tissue samples:

Add CB0094M-ES according to the ratio of tissue weight (g): CB0094M-ES volume (mL) = 1:5~10 (recommended: weigh approximately 0.1 g of tissue and add 1 mL of CB0094M-ES), then homogenize in an ice bath. Centrifuge at 8000 × g for 10 min at 4 °C. Collect the supernatant and keep it on ice for analysis.

3)Serum (plasma) samples: Analyze directly.

3. Assay Procedure

1. Pipette 0.3 mL of CB0094M-A into a 1.5 mL centrifuge tube, then add 0.1 mL of the sample and mix well.

2. Incubate in a 95 °C water bath for 30 min (cap tightly to prevent moisture loss), cool in an ice bath, then centrifuge at 10,000 × g, 25 °C for 10 min.

3. Transfer 200 μL of the supernatant to a micro glass cuvette or 96-well plate, measure the absorbance at 532 nm and 600 nm, record as A532 and A600. ΔA = A532 − A600.

4. Calculation of MDA Content

a. The calculation formula for measurements using micro glass cuvettes:

1.Calculation of MDA content in serum (plasma)

MDA content (nmol/ mL) = [ ΔA×V1÷(ε×d)×109]÷V2 = 25.8×ΔA

2.Calculation of MDA Content in Bacteria, Cells, or Animal Tissues

(1) Based on protein concentration MDAcontent (nmol/ mg prot) = [ ΔA×V1÷(ε×d)×109]÷(Cpr×V2) = 25.8× ΔA ÷Cpr

(2) Based on sample mass MDA content (nmol/g fresh sample weight) = [ ΔA×V1÷(ε×d)×109]÷(W ×V2÷V3) = 25.8×ΔA ÷W

(3) Based on bacterial or cell density MDA content (nmol/104) = [ ΔA×V1÷(ε×d)×109]÷(500×V2÷V3) = 0.0516×ΔA

Note：

V1：total reaction volume， 4×10-4 L；

ε：molar extinction coefficient of malondialdehyde (MDA)，155×103 L/mol/cm；

d：optical path length of the cuvette，1cm；

V2：volume of sample added，0.1 mL；

V3：volume of extraction solution added，1 mL；

Cpr：sample protein concentration，mg/mL；

W：sample mass，g；

500：total number of cells or bacteria，500 million.

b. The calculation formula for measurements using a 96-well plate:

1.Calculation of MDA content in serum (plasma)

MDA content (nmol/ mL) = [ ΔA×V1÷(ε×d)×109]÷V2 = 51.6×ΔA

2.Calculation of MDA Content in Bacteria, Cells, or Animal Tissues

(1) Based on protein concentration MDA content (nmol/ mg prot) = [ ΔA×V1÷(ε×d)×109]÷(Cpr×V2) = 51.6×ΔA÷Cpr

(2) Based on sample mass MDA content (nmol/g fresh sample weight) = [ ΔA×V1÷(ε×d)×109]÷(W ×V2÷V3) = 51.6×ΔA ÷W

(3) Base on bacterial or cell density MDA content (nmol/104) = [ ΔA×V1÷(ε×d)×109]÷(500×V2÷V3) = 0.1032×ΔA

Note:

V1：total reaction volume，4×10-4 L；

ε：molar extinction coefficient of malondialdehyde (MDA)，155×103 L/mol/cm；

d：Optical path length of a 96-well plate，0.5cm；

V2：volume of sample added，0.1 mL；

V3：volume of extraction solution added，1 mL；

Cpr：sample protein concentration，mg/mL；

W：sample mass，g；

500：total number of cells or bacteria，500 million.

1.For protein quantification, it is recommended to use BCA Protein Quantification Kit (C0050) .

2.The product is for R&D use only, not for diagnostic procedures, food, drug, household or other uses.

3.Please wear a lab coat and disposable gloves.

• CB0094M-Instruction Manual(285 KB)