QuicKey Human ALB(Albumin) ELISA Kit

SKU: E-TSEL-H0029

$ 331⁹⁵

QuicKey Human ALB(Albumin) ELISA Kit

Detection Range	1.56-100 ng/mL
Sensitivity	0.30 ng/mL

Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T
Assay time	2.5h
Detection range	1.56-100 ng/mL
Sensitivity	0.30 ng/mL
Sample type &Sample volume	serum and plasma; 50μL
Specificity	This kit recognizes Human ALB in samples. No significant or interference between Human ALB and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human ALB concentrations in serum and plasma.Please consult technical support for the applicability if other biological fluids need to be tested.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human ALB. Samples (or Standards) and biotinylated detection antibody specific for Human ALB are added to the micro ELISA plate wells. Human ALB would combine with the specific antibody. Then Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human ALB, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 ± 2 nm. The OD value is proportional to the concentration of Human ALB. You can calculate the concentration of Human ALB in the samples by comparing the OD of the samples to the standard curve.

Kit Components and Storage

An unopened kit can be stored at 2-8℃ for six months. After test, the unused wells and reagents should be stored according to the table below.

Item	Specifications	Storage conditions after test
Micro ELISA Plate (Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	2-8℃, 1 month
Reference Standard	96T: 2 vials 48T: 1 vial	Discard unused reconstituted standard and dilutions
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8℃
Biotinylated Detection Ab Working Solution	1 vial, 6 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	2-8℃ (Protect from light)
Substrate Reagent	1 vial, 10 mL	2-8℃ (Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Technical Data:

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human ALB were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ALB were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	4.06	10.37	43.36	4.47	10.60	44.40
Standard deviation	0.24	0.48	1.99	0.25	0.52	1.84
CV (%)	5.91	4.63	4.59	5.59	4.91	4.14

Recovery

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

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	87-101	95
EDTA plasma(n=8)	94-108	102

Target Information

Background

ALB is the most abundant protein in human plasma and serum. This protein is normally present in serum at concentrations ranging from 30 to 50 mg/mL and accounts for approximately 60% of the total protein content in serum [1]. ALB has a molecular weight of 66.7 kDa and is composed of a single polypeptide chain with 585 amino acids and 17 disulfide bonds. ALB is involved in many physiological processes. For instance, this protein aids in the regulation of osmotic pressure and pH in blood. In addition, ALB mediates lipid metabolism, sequesters toxins, and works as an antioxidant (i.e., it binds free radicals) [2]. Another important function of ALB is its action as a transport protein for a wide range of solutes that include some low mass hormones, fatty acids, and drugs
1. Hassan M, Azzazy E, Christenson R H. All About Albumin: Biochemistry, Genetics, and Medical Applications. Theodore Peters, Jr. San Diego, CA: Academic Press, 1996, 432 pp, $85.00. ISBN 0-12-552110-3[J]. 1997.
2. Otagiri M. A molecular functional study on the interactions of drugs with plasma proteins [J]. Drug metabolism and pharmacokinetics, 2005, 20(5): 309-323.

Database Links	SwissProt: P02768 Entrez Gene: 213
Synonyms	Serum albumin
Research Area	Cancer, Cardiovascular, Metabolism, Developmental biology, Signal transduction, Stem cells

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 90 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results