Human SEMA3B(Semaphorin 3B) ELISA Kit

Mfr.No.

E-EL-H0565

Antigen

SEMA3B

Alternative Names

LUCA-1 ; SEMA5 ; SEMAA ; SemA ; semaV

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 Human GP4/CD36 .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 Human GP4/CD36 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 Human GP4/CD36 ,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 Human GP4/CD36 . The concentration of Human GP4/CD36 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(ng/mL)	O.D	Average	Corrected
100	2.557 2.573	2.565	2.498
50	1.61 1.628	1.619	1.552
25	1.022 1.018	1.02	0.953
12.5	0.504 0.53	0.517	0.45
6.25	0.256 0.25	0.253	0.186
3.13	0.182 0.162	0.172	0.105
1.56	0.115 0.127	0.121	0.054
0	0.063 0.07099999999999999	0.067	--

Specificity

This kit recognizes Human SEMA3B in samples. No significant cross-reactivity or interference between Human SEMA3B 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

1.563--100ng/mL

Linearity

Samples were spiked with high concentrations of Human SEMA3B 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 (%)	93-106	98-114	92-106
1:2	Average (%)	99	104	100
1:4	Range (%)	92-103	82-96	89-104
1:4	Average (%)	98	88	95
1:8	Range (%)	88-100	80-93	81-97
1:8	Average (%)	95	85	88
1:16	Range (%)	94-108	81-94	87-99
1:16	Average (%)	100	86	93

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human SEMA3B were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human SEMA3B 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)	5.07	13.79	35.28	5.16	14.08	35.25
Standard	deviation	0.29	0.76	1.30	0.26	0.71	1.22
C V (%)	5.72	5.51	3.68	5.04	5.04	3.46

Recovery

The recovery of Human SEMA3B 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)	93-106	99
EDTA plasma (n=5)	92-103	98
Cell culture media (n=5)	86-100	91

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

0.94 ng/mL

Uniprot	Q13214

Manual

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SKU	Supplier No.	Size	Your price
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Human SEMA3B(Semaphorin 3B) ELISA Kit

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