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

Citations(16)Uniprot : P04924

Cat.No.:E-EL-RB0011
Reactivity: Rabbit

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Rabbit TNF-α in samples. No significant cross-reactivity or interference between Rabbit 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 Rabbit 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 Rabbit 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 Rabbit 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 Rabbit 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 Rabbit TNF-α. You can calculate the concentration of Rabbit 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 Rabbit 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 Rabbit 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)	49.73	154.44	395.12	47.58	159.52	432.48
Standard deviation	2.85	6.49	13.00	2.45	8.87	22.79
CV (%)	5.73	4.20	3.29	5.15	5.56	5.27

Recovery

The recovery of Rabbit 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)	96-109	101
EDTA plasma (n=8)	92-109	99
Cell culture media (n=8)	88-100	93

Linearity

Samples were spiked with high concentrations of Rabbit TNF-α and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P04924
Synonyms	DIF, TNF-alpha, TNFA, TNFSF2
Research Area	Cancer, Cardiovascular, 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

Vaccines (2020) IF: 4.086
An Antibody Persistent and Protective Two rSsCLP-Based Subunit Cocktail Vaccine against Sarcoptes scabiei in a Rabbit Model

DOI: 10.3390/vaccines8010129

PMID: 32187979
Sample: Serum
ARTIFICIAL ORGANS (2016) IF: 3.094
Hypothermic Machine Perfusion Reduced Inflammatory Reaction by Downregulating the Expression of Matrix Metalloproteinase 9 in a Reperfusion Model of Donation After Cardiac Death

DOI: 10.1111/aor.12658

Sample: Serum
Toxins (2018) IF: 3.273
Establishment of a New Zealand White Rabbit Model for Lethal Toxin (LT) Challenge and Efficacy of Monoclonal Antibody 5E11 in the LT-Challenged Rabbit Model

DOI: 10.3390/toxins10070289

Sample: Serum
World Journal of Stem Cells (2019) IF: 3.534
Anti-osteoarthritis effect of a combination treatment with human adipose tissue-derived mesenchymal stem cells and thrombospondin 2 in rabbits

DOI: 10.4252/wjsc.v11.i12.1115

PMID: 31875872
Sample: Synovia
Annals of Translational Medicine (2022) IF: 3.616
Seawater immersion exacerbates the pathological changes caused by incisive corneal injury in rabbit eyes

DOI: 10.21037/atm-22-2198

Sample: aqueous humor
Animals (2020) IF: 2.323
Inflammatory Correlated Response in Two Lines of Rabbit Selected Divergently for Litter Size Environmental Variability

DOI: 10.3390/ani10091540

PMID: 32882827
Sample: blood
INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE (2016) IF: 2.348
Hypothermic machine perfusion increases A20 expression which protects renal cells against ischemia/reperfusion injury by suppressing inflammation, apoptosis and necroptosis

DOI: 10.3892/ijmm.2016.2586

Sample: Tissue homogenate
MOLECULAR VISION (2017) IF: 2.367
Inhibitory effects of S100A4 gene silencing on alkali burn-induced corneal neovascularization: an in vivo study

PMID: 28479848
Sample: cornea
PLoS One (2020) IF: 2.74
Testicular toxicity following chronic codeine administration is via oxidative DNA damage and up-regulation of NO/TNF-α and caspase 3 activities

DOI: 10.1371/journal.pone.0224052

PMID: 32168344
Sample: Serum,Tissue homogenate
BIOMEDICINE & PHARMACOTHERAPY (2017) IF: 2.759
Oleanolic acid protects against pathogenesis of atherosclerosis, possibly via FXR-mediated angiotensin (Ang)-(1–7) upregulation

DOI: 10.1016/j.biopha.2017.11.151

Sample: Cell culture supernatant,Serum
MOLECULAR BIOLOGY REPORTS (2020) IF: 1.402
Codeine-induced hepatic injury is via oxido-inflammatory damage and caspase-3-mediated apoptosis

DOI: 10.1007/s11033-020-05983-6

PMID: 33211294
Sample: liver
CURRENT EYE RESEARCH (2019) IF: 1.672
The Efficiency of Cyclosporine A-Eluting Contact Lenses for the Treatment of Dry Eye

DOI: 10.1080/02713683.2018.1563702

PMID: 30580651
Sample: Tissue homogenate
Cutaneous and Ocular Toxicology (2021) IF: 1.82
L-carnosine mitigates interleukin-1α-induced dry eye disease in rabbits via its antioxidant, anti-inflammatory, antiapoptotic, and antifibrotic effects

DOI: 10.1080/15569527.2021.1935995

PMID: 34056995
Sample: serum
World Neurosurgery (2018) IF: 1.924
Neuroprotective Effects of Tocilizumab on Experimentally-Induced Spinal Cord Ischemia-Reperfusion Injury

DOI: 10.1016/j.wneu.2018.12.069

PMID: 30593966
Sample: Plasma,Tissue homogenate
THERIOGENOLOGY (2017) IF: 1.986
Effects of local lipopolysaccharide administration on the expression of Toll-like receptor 4 and pro-inflammatory cytokines in uterus and oviduct of rabbit does

DOI: 10.1016/j.theriogenology.2017.10.046

Sample: Plasma
JOURNAL OF OBSTETRICS AND GYNAECOLOGY (2018) IF: 0.545
Stress response to ovariohysterectomy in rabbits: role of anaesthesia and surgery

DOI: 10.1080/01443615.2017.1400523

Sample: Serum,Plasma

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