DESCRIPTION

MAX (MYC Associated Factor X) is a protein that plays a crucial role in the regulation of gene expression and cell cycle progression. MAX is a key partner of the MYC oncogene, forming heterodimers with MYC to regulate transcription through binding to E-boxes (5′-CACGTG-3′) in target gene promoters. MAX not only binds to MYC, but also interacts with other bHLH-ZIP family members (such as MAD and MNT) to form a complex regulatory network. The dysregulation of the MYC/MAX signaling pathway is closely related to the occurrence and development of various cancers. For example, overexpression of MYC is common in many cancers, and mutation or abnormal expression of MAX may also affect its ability to bind to MYC, thereby changing the gene regulatory network.

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	7.36	0.47	6.38
2	8	1.85	0.17	9.38
3	8	1.06	0.05	4.97

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	7.42	0.49	6.61
2	16	1.85	0.16	8.42
3	16	1.07	0.06	5.65

RECOVERY

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

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:32	101	98-102
	1:64	108	103-111

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	104
	Range (%)	101-107
1:8	Average% of Expected	90
	Range (%)	87-93
1:16	Average% of Expected	72
	Range (%)	70-73

CALIBRATION

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