DESCRIPTION

p63 is a member of the p53 family, which also includes p73. Like other members of the p53 family, the TP63 gene is expressed as multiple isoforms with distinct properties, including a full length and an amino-deleted isoform, named TAp63 andΔNp63, respectively. These isoforms have diverse functions and are involved in a wide range of biological processes. p63 plays a crucial role in the development and homeostasis of epithelial tissues. It regulates genes associated with cell survival, stem cell selfrenewal, migration, differentiation, and epithelial-to-mesenchymal transition. Specifically, p63 is essential for the development of epithelial tissues such as the epidermis, mammary glands, and other stratified epithelia. In pathological conditions, p63 is commonly expressed in normal tissues of epithelial origin and in lesions derived from these tissues.

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 (ng/mL)	SD	CV%
1	8	5.83	0.04	0.69
2	8	1.52	0.03	1.97
3	8	0.79	0.02	2.53

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	5.89	0.08	1.36
2	16	1.52	0.03	1.97
3	16	0.80	0.05	6.25

RECOVERY

The recovery of human p63 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	109	104-115
	1:64	111	108-115

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	102
	Range (%)	95-109
1:8	Average% of Expected	100
	Range (%)	87-114
1:16	Average% of Expected	98
	Range (%)	94-102

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human p63 One-Step ELISA Kit produced at Sunlong Medical™.