DESCRIPTION

Beta-2-microglobulin (B2M) is a 12-kDa protein comprised of 99 amino acids. B2M is a component of MHC class I molecules, which are present on the surface of nearly all nucleated cells. It can be found in body fluids under physiologic conditions as a result of shedding from cell surfaces or intracellular release. B2M has various biological functions, including antigen presentation. Serum B2M is regarded as a marker of disease severity in renal injury, infections, amyloidosis, and aging-related diseases. Investigations reveal that increased synthesis and release of B2M are present in several malignant diseases, including multiple myeloma, lymphoma, and solid tumors.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the solution yellow. The color intensity is proportional to the quantity of bound protein, which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	514.7	10.3	2.0
2	8	130.4	2.5	1.9
3	8	68.3	1.0	1.5

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	509.6	15.8	3.1
2	16	132.2	6.6	5.0
3	16	69.1	1.6	2.3

RECOVERY

The recovery of mouse Beta-2-Microglobulin spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:400,000	95	93-99
	1:800,000	95	91-100
Cell culture supernatant	1:4,000	101	96-108
Urine	1:100	96	93-101
	1:200	98	89-103

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The mouse serum was initially diluted 1:50,000. The cell culture supernatant was initially diluted 1:500. The urine was initially diluted 1:400.)

		Mouse serum	Cell culture supernatant	Urine
1:2	Average% of Expected	100	100	100
	Range (%)	-	-	-
1:4	Average% of Expected	110	88	92
	Range (%)	101-119	78-97	90-94
1:8	Average% of Expected	105	81	91
	Range (%)	96-114	78-84	90-92
1:16	Average% of Expected	84	80	75
	Range (%)	83-86	76-84	71-79

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant mouse Beta-2-Microglobulinin produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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• Price: USD, per kit (48T / 96T options available)