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Porcine TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit

Citations(8)Uniprot : P23563

Cat.No.:E-EL-P0010
Reactivity: Porcine

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T
Assay time	4.5h
Detection range	31.25-2000 pg/mL
Sensitivity	18.75 pg/mL
Sample type &Sample volume	Serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Porcine TNF-α in samples. No significant cross-reactivity or interference between Porcine TNF-α 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 Porcine TNF-α 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 Porcine TNF-α. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Porcine TNF-α 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 Porcine TNF-α, 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 Porcine TNF-α. You can calculate the concentration of Porcine TNF-α 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 Porcine TNF-α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Porcine TNF-α 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 (pg/mL)	92.68	184.99	768.41	87.57	181.59	820.00
Standard deviation	5.57	9.23	27.43	4.52	7.43	38.54
CV (%)	6.01	4.99	3.57	5.16	4.09	4.70

Recovery

The recovery of Porcine TNF-α 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)	96-108	101
EDTA plasma (n=8)	86-98	91
Cell culture media (n=8)	84-96	90

Target Information

Database Links	SwissProt: P23563
Synonyms	DIF, TNF-alpha, TNFA, TNFSF2

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

Journal of Translational Medicine (2022) IF: 8.44
Clinical improvement of sepsis by extracorporeal centrifugal leukocyte apheresis in a porcine model

DOI: 10.1186/s12967-022-03752-6

Sample: plasma
Animal Nutrition (2021) IF: 6.383
Dietary ferulic acid and vanillic acid on inflammation, gut barrier function and growth performance in lipopolysaccharide-challenged piglets

DOI: 10.1016/j.aninu.2021.06.009

Sample: serum
Animal Nutrition (2022) IF: 6.383
Dietary dimethylglycine sodium salt supplementation alleviates redox status imbalance and intestinal dysfunction in weaned piglets with intrauterine growth restriction

DOI: 10.1016/j.aninu.2021.12.007

PMID: 35785256
Sample: jejunum
Journal of Animal Science and Biotechnology (2020) IF: 4.167
Dietary protocatechuic acid ameliorates inflammation and up-regulates intestinal tight junction proteins by modulating gut microbiota in LPS-challenged piglets

DOI: 10.1186/s40104-020-00492-9

PMID: 32944233
Sample: Serum
MOLECULES (2019) IF: 3.06
Curcumin and Resveratrol Regulate Intestinal Bacteria and Alleviate Intestinal Inflammation in Weaned Piglets

DOI: 10.3390/molecules24071220

PMID: 30925757
Sample: Serum,Tissue homogenate
AMERICAN JOURNAL OF CHINESE MEDICINE (2020) IF: 3.682
Koumine Alleviates Lipopolysaccharide-Induced Intestinal Barrier Dysfunction in IPEC-J2 Cells by Regulating Nrf2/NF-κB Pathway

DOI: 10.1142/S0192415X2050007X

Sample: Cell culture medium
Biomedical Materials (2021) IF: 3.715
Preclinical efficacy study of a porous biopolymeric scaffold based on gelatin-hyaluronic acid-chondroitin sulfate in a porcine burn injury model: role of critical molecular markers (VEGFA, N-cadherin, COX-2), gamma sterilization efficacy and a comparison

DOI: 10.1088/1748-605X/ac1d3e

Sample: skin
Animals (2019) IF: 1.832
Effects of Maternal Supplementation with Rare Earth Elements during Late Gestation and Lactation on Performances, Health, and Fecal Microbiota of the Sows and Their Offspring

DOI: 10.3390/ani9100738

PMID: 31569383
Sample: Plasma,Feces

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