DESCRIPTION

PFKFB2(6-Phosphofructo-2-kinase/fructose 2,6-bisphosphatase 2) is also named as PFK-2/FBPase-2, which is a bifunctional enzyme responsible for the synthesis and breakdown of Fru-2,6-P2 in the regulation of glycolysis. PFKFB2 regulates glucose metabolism by controlling the level of fructose-2,6-bisphosphate. In the C-terminal section, it belongs to the phosphoglycerate mutase family. The enzyme is a dimer of identical subunits, each composed of 505 amino acids and containing three discrete domains. It has 2 isoforms produced by alternative splicing.

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns 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 (ng/mL)	SD	CV%
1	8	11.26	0.50	4.45
2	8	2.70	0.08	3.01
3	8	1.46	0.03	1.75

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	10.85	0.63	5.77
2	16	2.69	0.06	2.25
3	16	1.41	0.06	4.23

RECOVERY

The recovery of human PFKFB2 spiked to three different levels throughout the range of the assay in cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:16	98	94-104
	1:32	99	88-108

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 cell lysate was initially diluted 1:4.)

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	107
	Range (%)	102-113
1:8	Average% of Expected	76
	Range (%)	72-81
1:16	Average% of Expected	114
	Range (%)	104-123

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human PFKFB2 Sandwich ELISA Kit produced at Sunlong Medical™.

Ordering & shipping

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