Application

This kit can be used to measure the Albumin (ALB) content in serum, plasma samples.

Detection significance

Albumin is the most abundant plasma protein, with a molecular weight of 66.5 kDa. Albumin is synthesized in the liver at a rate of 9 to 12 grams per day and regulated by insulin, amino acid intake and low colloidal osmotic pressure. Changes of albumin content in urine and serum are predictors of diabetic nephropathy, cardiovascular disease, liver disease and sepsis.

Detection principle

Bromocresol green (BCG) is widely used as protein staining agent. BCG can combine the albumin in pH 4.0~4.2 to form an albumin-BCG complex. And the color changed from yellow to green. The depth of color is proportional to the concentration of albumin. The content of albumin in serum can be calculated indirectly by measuring the OD value at 630 nm.

Experimental instrument

Micropipette, Vortex mixer, Centrifuge, Microplate reader (630 nm)

Sample preparation

Serum (plasma) sample: detect directly.

Operation steps

It is recommended to take 2~3 samples which expected large difference to do pre-experiment before formal experiment. According to the results of pre-experiment and the linear range (0.08-15 g/L) of the kit, then dilute the samples.

1. Dilution of standard

Dilute 20 g/L standard solution with double distilled water to a serial concentration. The recommended dilution gradient is as follows: 15, 12, 8, 5, 3.5, 2, 1, 0 μg/mL.

2. Assay steps:

(1) Standard well: add 10 μL standard with different concentration into the wells.

Sample tube: add 10 μL of sample into the wells.   

(2) Add 250 μL of the Reagent 1 working solution to each well.

(3) Stand for 10 min at room temperature.

(4) Measure the OD value of each well at 630 nm with Microplate reader.

Note: It can be refer to the following operating table.

Technical parameters

1. The sensitivity of the kit is 0.08 g/L.

2. The intra-assay CV is 1.5 % and the inter-assay CV is 4.6 %.

3. The recovery of the kit is 95 %.

4. The linear range of the kit is 0.08 g/L-15 g/L.

Notes

1. The kit is for scientific research only.

2. Instructions should be followed strictly, changes of operation may result in unreliable results.

3. The validity of kit is 6 months.

4. Do not use components from different batches of kit.