DESCRIPTION

PSMD14, also known as POH1, is a highly conserved protein in eukaryotes, playing a crucial role in cellular homeostasis. It is a subunit of the regulatory particle of the proteasome and functions as an intrinsic deubiquitinase. Specifically, PSMD14 removes polyubiquitin chains from substrate proteins, which is essential for the translocation of substrates into the core of the proteasome for degradation. Moreover, inhibitors targeting PSMD14 have shown promising anticancer activity in cell lines, indicating its potential as a druggable target.

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	5.57	0.26	4.74
2	8	1.41	0.05	3.29
3	8	0.66	0.03	4.93

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	5.66	0.25	4.37
2	16	1.44	0.07	4.99
3	16	0.69	0.04	6.33

RECOVERY

The recovery of human PSMD14/POH1 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	88	83-93

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 1:2.)

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	89
	Range (%)	85-94
1:8	Average% of Expected	96
	Range (%)	84-108
1:16	Average% of Expected	104
	Range (%)	89-119

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human PSMD14/POH1 Sandwich ELISA Kit produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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