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Human C3a(Complement Component 3a) ELISA Kit

Citations(5)Uniprot : P01024

Cat.No.:E-EL-H0818
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.31-20 ng/mL
Sensitivity	0.19 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human C3a in samples. No significant cross-reactivity or interference between Human C3a 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 C3a 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 C3a. 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 C3a 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 C3a, 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 C3a. You can calculate the concentration of Human C3a 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 C3a were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human C3a 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)	1.01	2.09	8.39	0.94	2.22	7.83
Standard deviation	0.07	0.09	0.30	0.05	0.13	0.24
CV (%)	6.93	4.31	3.58	5.32	5.86	3.07

Recovery

The recovery of Human C3a 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)	84-97	90
EDTA plasma (n=8)	91-102	97
Cell culture media (n=8)	87-98	93

Linearity

Samples were spiked with high concentrations of Human C3a and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P01024
Synonyms	C3a-desArg, ASP
Research Area	Cancer, Immunology, Signal transduction

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

Frontiers in Immunology (2022) IF: 8.786
Immune modulation as a consequence of SARS-CoV-2 infection.

DOI: 10.3389/fimmu.2022.954391

Sample: serum
Frontiers in Immunology (2020) IF: 5.085
Complement Expression and Activation in Osteoarthritis Joint Compartments

DOI: 10.3389/fimmu.2020.535010

PMID: 33193305
Sample: Cell culture supernatant
Scientific Reports (2022) IF: 4.38
Brief report on the relation between complement C3a and anti dsDNA antibody in systemic lupus erythematosus

DOI: 10.1038/s41598-022-10936-z

Sample: serum
PLoS Neglected Tropical Diseases (2021) IF: 4.411
Anaphylatoxins orchestrate Th17 response via interactions between CD16+ monocytes and pleural mesothelial cells in tuberculous pleural effusion

DOI: 10.1371/journal.pntd.0009508

PMID: 34237073
Sample: pleural fluid,PMCs cells,monocytes
REACTIVE & FUNCTIONAL POLYMERS (2021) IF: 3.975
Antioxidant and antithrombotic study of novel chitosan-diallyl disulfide inclusion complexes nanoparticles for hemodialysis applications

DOI: 10.1016/j.reactfunctpolym.2021.104894

Sample: Plasma

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