Human LFA-3/CD58(Lymphocyte Function Associated Antigen 3) ELISA Kit

Mfr.No.

E-EL-H1689

Antigen

LFA-3/CD58

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 DMD .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 DMD 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 DMD ,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 DMD . The concentration of Mouse DMD 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
800	2.474 2.496	2.485	2.416
400	1.617 1.639	1.628	1.559
200	0.931 0.917	0.9239999999999999	0.855
100	0.465 0.483	0.474	0.405
50	0.273 0.257	0.265	0.196
25	0.182 0.162	0.172	0.103
12.50	0.112 0.132	0.122	0.053
0	0.062 0.076	0.06900000000000001	--

Specificity

This kit recognizes Human LFA-3/CD58 in samples. No significant cross-reactivity or interference between Human LFA-3/CD58 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

12.5--800pg/mL

Linearity

Samples were spiked with high concentrations of Human LFA-3/CD58 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 (%)	93-108	88-103	88-101
1:2	Average (%)	100	94	96
1:4	Range (%)	90-103	84-95	82-92
1:4	Average (%)	97	90	87
1:8	Range (%)	90-103	84-95	88-100
1:8	Average (%)	95	89	94
1:16	Range (%)	89-100	83-96	87-100
1:16	Average (%)	94	88	93

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human LFA-3/CD58 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human LFA-3/CD58 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)	41.93	77.50	381.68	45.46	78.71	413.67
Standard	deviation	2.53	3.49	14.62	2.59	4.36	16.80
C V (%)	6.03	4.50	3.83	5.70	5.54	4.06

Recovery

The recovery of Human LFA-3/CD58 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)	88-99	94
EDTA plasma (n=5)	88-102	95
Cell culture media (n=5)	87-100	94

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

7.5 pg/mL

Uniprot	P19256

Manual

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SKU	Supplier No.	Size	Your price
BHE10304058	E-EL-H1689	96T	$609	Add to Cart
BHE10304058	E-EL-H1689	96T*5	$2740.50	Add to Cart
BHE10304058	E-EL-H1689	96T*10	$5176.50	Add to Cart
BHE10304058	E-EL-H1689	48T	$487.20	Add to Cart

Human LFA-3/CD58(Lymphocyte Function Associated Antigen 3) ELISA Kit

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