DESCRIPTION

EH domain-containing protein 2 (EHD2) is a member of the dynamin-related ATPase family that orchestrates membrane remodeling and vesicular trafficking at the plasma membrane. First identified as a caveolar-associated protein, EHD2 forms ATPdependent oligomers that tether caveolae to the underlying actin cytoskeleton, thereby stabilizing these invaginated microdomains and restraining their internalization. Beyond its membrane-sculpting role, EHD2 also shuttles to the nucleus where it functions as a transcriptional repressor, linking mechanical cues sensed at caveolae to gene-expression programs. In adipocytes, EHD2 is enriched at lipid-droplet contact sites and is required for insulin-dependent GLUT4 translocation, highlighting its importance for glucose homeostasis. Decreased EHD2 expression has been associated with poor prognosis in hepatocellular carcinoma and breast cancer, suggesting a tumor-suppressive function. Thus, EHD2 operates as a mechanotransducer that integrates membrane dynamics, metabolic signaling, and transcriptional control.

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	8	11.76	0.22
2	8	8	3.23	0.09
3	8	8	1.43	0.04

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	12.47	0.84	6.70
2	16	3.25	0.10	3.09
3	16	1.52	0.10	6.83

RECOVERY

The recovery of human/mouse EHD2 spiked to three different levels throughout the range of the assay in cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:160	115	108-126

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 1:20. )

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	117
	Range (%)	113-121
1:8	Average% of Expected	91
	Range (%)	85-98
1:16	Average% of Expected	111
	Range (%)	100-122

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human/mouse EHD2 produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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