Human Na+ Taurocholate Cotransporting Polypeptide (NTCP) ELISA Kit

Principle of the Assay

The microtiter plate provided in this kit has been pre-coated with an antibody specific to NTCP. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to NTCP. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain NTCP, biotin-conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution, and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of NTCP in the samples is then determined by comparing the O.D. of the samples to the standard curve.

For use with: Tissue homogenates, Cell lysates, and Other biological fluids.

For Research Use Only. Not for use in diagnostic procedures.

Target Information

As a major transporter of conjugated bile salts from plasma into the hepatocyte, it has a key role in the enterohepatic circulation of bile salts necessary for the solubilization and absorption of dietary fat and fat-soluble vitamins. It exhibits broad substrate specificity and transports various non-bile acid organic compounds as well. It is strictly dependent on the extracellular presence of sodium. Able to transport taurocholate, cholate, and the non-bile acid estron sulfate (PubMed:14660639, PubMed:24867799).

GENE ID	6554
SWISS PROT	Q14973
SYNONYMS	SLC10A1; NTCP1; Solute Carrier Family 10 Member 1,Sodium/Bile Acid Cotransporter Family; Cell growth-inhibiting gene 29 protein; Na(+)/bile acid cotransporter

Materials Supplied

Kit Components	96 Wells Quantity/Size
Pre-coated, ready-to-use 96-well strip plate	1 plate
Plate sealer for 96 wells	2
Standard	2 tubes
Diluent buffer	1 bottle
Detection Reagent A	1 bottle
Detection Reagent B	1 bottle
TMB Substrate	1 tube
Stop Solution	1 tube
Wash Buffer (30X concentrate)	1 tube
Product data sheet	1 copy

Storage

Storage

The TMB Substrate, Wash Buffer (30X concentrate), and the Stop Solution should be stored at 4°C upon receipt, while the other items should be stored at -20°C.

Performance Characteristics

REPEATABILITY	Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level NTCP were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle, and high-level NTCP were tested on 3 different plates, with 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
SENSITIVITY	The minimum detectable dose was 0.27ng/mL.
ASSAY RANGE	0.781–50ng/mL
SPECIFICITY	This assay has high sensitivity and excellent specificity for the detection of NTCP. No significant cross-reactivity or interference between NTCP and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between NTCP and all analogs, therefore, cross reactivity may still exist.