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Human SEMA3A(Semaphorin 3A) ELISA Kit

  • Cat.No.:E-EL-H1274

  • Reactivity: Human

To Purchase E-EL-H1274

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $609
Qty:

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.
ItemSpecificationsStorage
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 LinksSwissProt: Q14563
SynonymsSEMA3A, COLL1, HH16, Hsema-I, Hsema-III, SEMA1, SEMAD, SEMAIII, SEMAL, SemD, coll-1, semaphorin 3A

Assay Procedures

elisa assay procedure 1

1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C

elisa assay procedure 2

2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C

elisa assay procedure 3

3. Aspirate and wash the plate for 3 times

elisa assay procedure 4

4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 5

5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 6

6. Add 50μL Stop Solution

elisa assay procedure 7

7. Read the plate at 450nm immediately. Calculation of the results

Related Products

Elabscience® provides ancillary reagents required for ELISA kit developments.11 reagents in total, including reagents for coating, blocking, washing, improving the stability and reducing matrix interference etc.
Cat.No. Product Name Specifications
E-ELIR-001 ELISA Plate Coating Buffer(1×) 100mL, 1000mL
E-ELIR-002 ELISA Plate Coating Buffer(5×) 100mL, 1000mL
E-ELIR-003 ELISA Plate Blocking Buffer 100mL, 1000mL
E-ELIR-004 Wash Buffer for Sandwich-ELISA(25×) 100mL, 1000mL
E-ELIR-005 Wash Buffer for Competitive-ELISA(25×) 100mL, 1000mL
E-ELIR-006 Stop Solution 100mL, 1000mL
E-ELIR-007 HRP-conjugate Stabilizer 10mL, 100mL, 500mL
E-ELIR-008 HRP-conjugate Diluent 100mL, 1000mL
E-ELIR-009 Biotinylated Antibody Stabilizer 10mL, 100mL, 500mL
E-ELIR-0010 Biotinylated Antibody Diluent 10mL, 100mL, 500mL
E-ELIR-0011 Sample Diluent 10mL, 100mL, 500mL

Citations

From now on, if you have published a paper by using any of our products since 1/1/2019, fill out the “Elabscience Publication Reward Application Form”carefully and send it to orders@elabscience.com, we will get back to you with the reward after we confirm it ASAP!

View more details about our Publication Reward >>

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

    DOI: 10.1038/s41413-020-0096-1

    PMID: 32435517

    Sample: Cell culture medium
  2. Oncotarget (2016) IF: 5.168
    Axon Guidance Molecule Semaphorin3A Is A Novel Tumor Suppressor In Head And Neck Squamous Cell Carcinoma.

    DOI: 10.18632/oncotarget.6831

    PMID: 26755661

    Sample: Cell culture supernatant
  3. Journal of Biomedical Materials Research Part B-Applied Biomaterials (2016) IF: 3.189
    SEMA3A Suspended In Matrigel Improves Titanium Implant Fixation In Ovariectomized Rats.

    DOI: 10.1002/jbm.b.33730

    PMID: 27403923

    Sample: Others
  4. 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

    PMID: 28188869

    Sample: Serum
  5. Iranian Journal of Medical Sciences (2019) IF: 1.854
    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
  6. The Anatomical Record (2018)
    The Neuropilin-1 Ligand, Sema3A, Acts as a Tumor Suppressor in the Pathogenesis of Acute Leukemia

    DOI: 10.1002/ar.24016

    PMID: 30378769

    Sample: Plasma
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