Human L1CAM/CD171(L1-Cell Adhesion Molecule) ELISA Kit

Mfr.No.

E-EL-H1895

Antigen

L1CAM/CD171

Background

This kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse IgG2a .Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse IgG2a and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse IgG2a ,biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The OD value is measured spectrophotometrically at a wavelength of 450nm. The OD value is positively associated with the concentration of Mouse IgG2a . The concentration of Mouse IgG2a in the samples can be calculated by comparing the OD of the samples to the standard curve.

Description

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration(pg/mL)	O.D	Average	Corrected
6000	2.53 2.53	2.53	2.465
3000	1.607 1.641	1.624	1.559
1500	0.961 0.953	0.9570000000000001	0.892
750	0.514 0.534	0.524	0.459
375	0.27 0.25	0.26	0.195
187.5	0.18 0.154	0.167	0.102
93.75	0.11 0.124	0.117	0.052
0	0.06 0.07000000000000001	0.065	--

Specificity

This kit recognizes Human L1CAM/CD171 in samples. No significant cross-reactivity or interference between Human L1CAM/CD171 and analogues was observed.

Assay Time

4.5h

Components

An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	8 wells ×12 strips	-20℃, 6 months
Reference Standard	2 vials
Concentrated Biotinylated Detection Ab (100×)	1 vial, 120 μL
Concentrated HRP Conjugate (100×)	1 vial, 120 μL	-20℃(shading light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	4℃, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	4℃(shading light)
Stop Solution	1 vial, 10 mL	4℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Detection Method

Colormetric

Detection Range

93.75--6000pg/mL

Linearity

Samples were spiked with high concentrations of Human L1CAM/CD171 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	97-112	99-114	92-102
1:2	Average (%)	104	106	97
1:4	Range (%)	85-97	82-93	86-100
1:4	Average (%)	92	87	93
1:8	Range (%)	93-107	81-92	88-100
1:8	Average (%)	98	87	94
1:16	Range (%)	90-102	81-95	81-95
1:16	Average (%)	95	88	87

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human L1CAM/CD171 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human L1CAM/CD171 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)	275.61	768.72	2327.60	272.05	797.21	2387.94
Standard	deviation	17.97	34.36	125.92	15.51	33.96	90.03
C V (%)	6.52	4.47	5.41	5.70	4.26	3.77

Recovery

The recovery of Human L1CAM/CD171 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=5)	83-98	90
EDTA plasma (n=5)	88-101	93
Cell culture media (n=5)	93-107	99

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

56.25 pg/mL

Uniprot	P32004

Manual

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Human L1CAM/CD171(L1-Cell Adhesion Molecule) ELISA Kit

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