QuicKey Human TGF-α(Transforming Growth Factor Alpha) ELISA Kit

SKU: E-TSEL-H0027

$ 331⁹⁵
Save $ 59⁰⁰

QuicKey Human TGF-α(Transforming Growth Factor Alpha) ELISA Kit

Detection Range	31.25-2000 pg/mL
Sensitivity	7.54 pg/mL

Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T
Assay time	2.5h
Detection range	31.25-2000 pg/mL
Sensitivity	7.54 pg/mL
Sample type &Sample volume	serum and plasma; 50μL
Specificity	This kit recognizes Human TGF-α in samples. No significant or interference between Human TGF-α 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 TGF-α 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 TGF-α. Samples (or Standards) and biotinylated detection antibody specific for Human TGF-α are added to the micro ELISA plate wells. Human TGF-α 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 TGF-α, 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 TGF-α. You can calculate the concentration of Human TGF-α 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 TGF-α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TGF-α 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 (pg/mL)	88.62	231.38	839.43	89.90	213.36	948.10
Standard deviation	5.01	10.92	42.31	5.30	9.64	45.03
CV (%)	5.65	4.72	5.04	5.90	4.52	4.75

Recovery

The recovery of Human TGF-α 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)	101-110	105
EDTA plasma(n=8)	91-100	95

Target Information

Background

Transforming growth factor alpha (TGF-α) is a 50 amino acid protein encoded by the gene TGFA and it shares structural homology with Epidermal growth factor(EGF). As a member of the epidermal growth factor (EGF) family, TGF-α is a mitogenic polypeptide and becomes activated when binding to receptors capable of protein kinase activity for cellular signaling[1]. Although TGF-α is most often associated with its proliferation, differentiation, and transformation-promoting effects, it is also involved in angiogenesis, bone resorption, cell metabolism, cell migration and wound healing. Further, TGF-α has been implicated not only in myriad forms of cancer, but also in several neurodegenerative disorders[2].
1. Mead J E, Fausto N. Transforming growth factor alpha may be a physiological regulator of liver regeneration by means of an autocrine mechanism[J]. Proceedings of the National Academy of Sciences, 1989, 86(5): 1558-1562.
2. Junier M P. What role (s) for TGFα in the central nervous system?[J]. Progress in neurobiology, 2000, 62(5): 443-473.

Database Links	SwissProt: P01135 Entrez Gene: 7039
Synonyms	TGFA, ETGF
Research Area	Cancer, Cardiovascular, Signal transduction

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