DESCRIPTION

Kinesin superfamily proteins (KIFs) are microtubule-based molecular motors essential for the intracellular transport of various cargos, including organelles, proteins, and RNAs. KIF5A is expressed exclusively in neurons and transports neuronal cargoes into axons and dendrites. KIF5A mutations have been associated with Charcot-Marie-Tooth Type 2, an axonal peripheral neuropathy characterized by progressive loss of peripheral sensation and muscle wasting.

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	9.50	0.60	6.28
2	8	2.46	0.07	2.69
3	8	1.31	0.05	3.92

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	9.71	0.72	7.46
2	16	2.41	0.13	5.60
3	16	1.25	0.08	6.00

RECOVERY

The recovery of human/mouse KIF5A spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:32	96	89-103
	1:64	89	86-92
Tissue lysate	1:8	94	86-99

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 1:4.)

		Cell lysate(Sample Diluent PT 4)	Tissue lysate(Sample Diluent PT 4B1)
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	125	107
	Range (%)	117-130	104-109
1:8	Average% of Expected	98	97
	Range (%)	82-110	96-98
1:16	Average% of Expected	83	99
	Range (%)	76-103	98-100

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human/mouse KIF5A produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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• Price: USD, per kit (48T / 96T options available)