DESCRIPTION

Inhibitor of apoptosis-stimulating protein of p53 (iASPP), encoded by PPP1R13L gene, is often overexpressed in human cancers. The ASPP family includes three members, namely ASPP1, ASPP2, and iASPP, which are specific regulators of p53-, p63-, and p73mediated apoptosis. ASPP1 and ASPP2 enhance the apoptotic function of p53, whereas iASPP specifically inhibits p53-mediated apoptosis. Overexpression of iASPP is associated with resistance to cisplatin-induced apoptosis and rediation therapy. iASPP plays a pivotal role in regulating cancer cell proliferation and tumor progression.

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	3.18	0.11	3.46
2	8	0.82	0.02	2.44
3	8	0.45	0.02	4.44

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	2.91	0.30	10.31
2	16	0.74	0.09	12.16
3	16	0.42	0.04	9.52

RECOVERY

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

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:256	101	95-108
	1:512	107	106-108

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 samples were initially diluted 1:32.)

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	109
	Range (%)	108-111
1:8	Average% of Expected	112
	Range (%)	106-118
1:16	Average% of Expected	97
	Range (%)	94-101

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human iASPP 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)