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Rat C5a(Complement Component 5a) ELISA Kit

Cat.No.:E-EL-R3011
Reactivity: Rat

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	15.63-1000 ng/mL
Sensitivity	9.38 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Rat C5a in samples. No significant cross-reactivity or interference between Rat C5a 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 Rat C5a 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 Rat C5a. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat C5a 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 Rat C5a, 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 Rat C5a. You can calculate the concentration of Rat C5a 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 Rat C5a were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat C5a 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)	44.08	113.30	497.15	45.05	120.13	443.23
Standard deviation	2.46	5.35	26.40	2.58	5.93	22.69
CV (%)	5.58	4.72	5.31	5.73	4.94	5.12

Recovery

The recovery of Rat C5a 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)	91-103	97
EDTA plasma (n=8)	92-104	99
Cell culture media (n=8)	87-97	92

Linearity

Samples were spiked with high concentrations of Rat C5a and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Synonyms	C5a
Research Area	Cell Biology, Metabolism, 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

Computational and Structural Biotechnology Journal (2020) IF: 6.018

Lipid metabolic signatures deviate in sepsis survivors compared to non-survivors

DOI: 10.1016/j.csbj.2020.11.009
PMID: 33304464	Sample: plasma

Frontiers in Pharmacology (2022) IF: 5.811

Guhong Injection Prevents Ischemic Stroke-Induced Neuro-Inflammation and Neuron Loss Through Regulation of C5ar1.

DOI: 10.3389/fphar.2022.818245
	Sample: brain

SHOCK (2017) IF: 3.113

Tranexamic Acid Attenuates The Loss of Lung Barrier Function in a Rat Model of Polytrauma And Hemorrhage With Resuscitation

DOI: 10.1097/SHK.0000000000000758
	Sample: Lung tissue

NEUROTOXICITY RESEARCH (2019) IF: 3.911

The Novel C5aR Antagonist DF3016A Protects Neurons Against Ischemic Neuroinflammatory Injury

DOI: 10.1007/s12640-019-00026-w
PMID: 30953275	Sample: Cell culture supernatant

TRANSPLANT IMMUNOLOGY (2022) IF: 1.708

Effect of a C5a receptor antagonist on macrophage function in an intestinal transplant rat model

DOI: 10.1016/j.trim.2022.101559
PMID: 35227893	Sample: serum

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Reviews/Q&A

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Reviews
Q&A

C*************aSubmitted [ Nov 15 2021 ]

Asked: Hello, I am Chiyoshi Toyama from Japan. I purchased your product (Cat: E-EL-R3011) and plan to test C5a in rat serum. Do you have any data on the normal range of C5a in serum? Thank you for your kin. Sincerely,

adminSubmitted [ Nov 15 2021 ]

Answered: Dear Toyama san,thanks for your message.For the kit E-EL-R3011, we have validated with normal rat serum samples, the reference value of tested samples were about 20-300 ng/mL, with individual difference. We will send the test report to your email later, please check it,thank you. best regards,

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