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Human SEMA3A(Semaphorin 3A) ELISA Kit

Citations(11)Uniprot : Q14563

Cat.No.:E-EL-H1274
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.16-10 ng/mL
Sensitivity	0.10 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human SEMA3A in samples. No significant cross-reactivity or interference between Human SEMA3A 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 SEMA3A concentrations in serum, plasma and other biological fluids.

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 SEMA3A. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human SEMA3A and 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 SEMA3A, 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 nm ± 2 nm. The OD value is proportional to the concentration of Human SEMA3A. You can calculate the concentration of Human SEMA3A in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 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	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	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 SEMA3A were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human SEMA3A 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)	0.55	0.89	4.77	0.60	0.88	5.24
Standard deviation	0.03	0.05	0.18	0.03	0.04	0.22
CV (%)	5.45	5.62	3.77	5.00	4.55	4.20

Recovery

The recovery of Human SEMA3A 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)	92-108	99
EDTA plasma (n=8)	87-98	92
Cell culture media (n=8)	89-104	96

Linearity

Samples were spiked with high concentrations of Human SEMA3A and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: Q14563
Research Area	Cancer, Cardiovascular, Neuroscience

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

Bone Research (2020) IF: 11.508
Osteoblasts are inherently programmed to repel sensory innervation

DOI: 10.1038/s41413-020-0096-1

Sample: Cell culture medium
Oncotarget (2016) IF: 5.008
Axon guidance molecule semaphorin3A is a novel tumor suppressor in head and neck squamous cell carcinoma

DOI: 10.18632/oncotarget.6831

Sample: Cell culture supernatant
JOURNAL OF PERIODONTAL RESEARCH (2022) IF: 4.419
Gingival crevicular fluid lipocalin-2 and semaphorin3A in stage III periodontitis: Non-surgical periodontal treatment effects

DOI: 10.1111/jre.12995

PMID: 35468224
Sample: gingival crevicular fluid (GCF)
PLoS One (2021) IF: 3.24
Extract of Scutellaria baicalensis induces semaphorin 3A production in human epidermal keratinocytes

DOI: 10.1371/journal.pone.0250663

PMID: 33905439
Sample: Cell lysate
RHEUMATOLOGY INTERNATIONAL (2022) IF: 3.58
Assessment of semaphorin 3A and semaphorin 7A levels in primary Sjogren's syndrome

DOI: 10.1007/s00296-022-05197-y

PMID: 36030414
Sample: serum
INTERNATIONAL OPHTHALMOLOGY (2022) IF: 2.029
Evaluation of serum semaphorin 3A and interleukin 6 levels in patients with pseudoexfoliation syndrome

DOI: 10.1007/s10792-022-02599-6

Sample: serum
GENE (2017) IF: 2.415
Decreased expression of Sema3A, an immune modulator, in blood sample of multiple sclerosis patients

DOI: 10.1016/j.gene.2017.02.013

Sample: Serum
International Journal of Endocrinology (2022) IF: 2.803
Identification and Functional Characterization of a Novel Variant in the SEMA3A Gene in a Chinese Family with Kallmann Syndrome

DOI: 10.1155/2022/2504660

Sample: COS-7 cells
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART B-APPLIED BIOMATERIALS (2016) IF: 2.881
SEMA3A suspended in matrigel improves titanium implant fixation in ovariectomized rats

DOI: 10.1002/jbm.b.33730

Sample: Others
Anatomical Record-Advances in Integrative Anatomy and Evolutionary Biology (2018) IF: 1.373
The Neuropilin‐1 Ligand, Sema3A, Acts as a Tumor Suppressor in the Pathogenesis of Acute Leukemia

DOI: 10.1002/ar.24016

Sample: Plasma
Iranian Journal of Medical Sciences (2019)
Negative Regulation of Semaphorin-3A Expression in Peripheral Blood Mononuclear Cells Using MicroRNA-497-5p

DOI: 10.30476/IJMS.2019.44955

PMID: 31439976
Sample: Cell culture supernatant

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