DESCRIPTION

Rho-associated protein kinase 1 (ROCK1) is a serine/threonine protein kinase that was identified as a RhoGTP-binding protein. To date, two isoforms encoded by two different genes of ROCK (ROCK1 and ROCK2) have been investigated. ROCK1 and ROCK2 have been shown to fulfill major roles in carcinogenesis. These two proteins share an overall sequence similarity in their kinase domains of 92%, and at the amino-acid level, a similarity of 65%. ROCK1 is involved in the pathogenesis of metabolic-related diseases, including hypertension, arteriosclerosis, Alzheimer's disease, and diabetes.

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 20 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	20	992.5	53.0	5.3
2	20	244.5	17.2	7.0
3	20	59.0	5.2	8.9

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	885.5	85.7	9.7
2	24	246.9	14.8	6.0
3	24	60.6	2.5	4.2

RECOVERY

The recovery of human ROCK1 spiked to three different levels throughout the range of the assay in cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:128	84	78-89
	1:256	80	78-82

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 1:16.)

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	108
	Range (%)	103-112
1:8	Average% of Expected	115
	Range (%)	114-116
1:16	Average% of Expected	120
	Range (%)	119-120

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human ROCK1 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)