| KEAP1

Kelch-like ECH-associated protein 1 is a protein that is encoded by the Keap1 gene.Keap1 has four discrete protein domains. The N-terminal Broad complex, Tramtrack and Bric-à-Brac (BTB) domain contains the Cys151 residue, which is one of the important cysteines in stress sensing. The intervening region (IVR) domain contains two critical cysteine residues, Cys273 and Cys288, which are a second group of cysteines important for stress sensing. A double glycine repeat (DGR) and C-terminal region domains collaborate to form a β-propeller structure, which is where Keap1 interacts with Nrf2.
 

| Typical data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, wash technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data are provided below for reference only.

 

Concentration (ng/mL)	10	5	2.5	1.25	0.625	0.312	0.156	0
OD	2.08	1.31	0.75	0.5	0.33	0.25	0.21	0.09
Corrected OD	1.99	1.22	0.66	0.41	0.24	0.16	0.12	-

 

| Repeatability

Intra-assay Precision (Precision within an assay)：CV% < 10%; Inter-assay Precision (Precision between assays)：CV% < 10%
 

| Recovery

The recovery of Human KEAP1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

 

Sample Type	Range(%)	Average Recovery(%)
Tissue Homogenates(n=8)	84-101	96
Cell Culture Supernates(n=8)	96-107	104

 

| Linearity

Samples were spiked with high concentrations of Human KEAP1 and diluted with Universal Diluent to produce samples with values within the range of the assay.

 

		Tissue Homogenates(n=4)	Cell Culture Supernates(n=4)
1 : 2	Range (%)	84-96	90-110
	Average (%)	92	96
1 : 4	Range (%)	89-103	105-115
	Average (%)	94	108