Size	1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Species	Rat
Accession Number	P16599
Gene Id	103694380\|24835
Gene Symbols	TNF\|TNFSF2
Protein Name / Synonyms	Tumor necrosis factor (Cachectin) (TNF-alpha) (Tumor necrosis factor ligand superfamily member 2) (TNF-a) [Cleaved into: Tumor necrosis factor, membrane form (N-terminal fragment) (NTF) Intracellular domain 1 (ICD1) Intracellular domain 2 (ICD2) C-domain 1 C-domain 2 Tumor necrosis factor, soluble form]
Quantitative/Semi-Quantitative	Quantitative
Specificity	This ELISA kit shows no cross-reactivity with any of the cytokines tested: rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3 alpha, beta- NGF, TIMP-1, VEGF.
Compatible Sample Types	Cell Culture Supernatants, Plasma, Serum, Tissue Lysates, Cell Lysates
Solid Support	96-well Microplate
Method Of Detection	Colorimetric
Design Principle	Sandwich-based
Sensitivity	25 pg/ml Need more sensitivity? Check out the new BIQ-ELISA™ kit for this target. Still not enough? Then your answer is our Ultrasensitive Biomarker Testing Service powered by Simoa^™ technology.
Detection Range	25 pg/ml - 20000 pg/ml
Recommended Dilution (Serum/Plasma)	2 fold
Estimated Lead Time	1-2 business days
Shipping Type	Blue ice
Storage	≤-20°C

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Target Background

Tumor necrosis factor alpha (TNF alpha), also known as lymphotoxin and cachectin, is a pleiotropic cytokine. TNF alpha is a smaller cytokine with a molecular weight of 26 kDa (transmembrane) and 17 kDa (soluble) that binds to the same receptors producing a vast array of effects similar to those of TNF-beta. Both TNF-alpha and TNF-beta share 30% amino acid homology and have similar biological activities. TNF-alpha is produced primarily by monocytes, neutrophils and macrophages. It is found in synovial cells and macrophages in the tissues. TNF-alpha and TNF-beta can modulate many immune and inflammatory functions, while having the ability to inhibit tumor growth. Target tumor cells must express TNF receptors 1 (55 kDa) and 2 (75 kDa) to be killed, with the p55 receptor mediating the cytotoxic response.

Product Features

Additional Notes

Strip plates and additional reagents allow for use in multiple experiments
Quantitative protein detection
Establishes normal range
The best products for confirmation of antibody array data

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components

Pre-Coated 96-well Strip Microplate
Wash Buffer
Stop Solution
Assay Diluent(s)
Lyophilized Standard
Biotinylated Detection Antibody
Streptavidin-Conjugated HRP
TMB One-Step Substrate

Other Materials Required

Distilled or deionized water
Precision pipettes to deliver 2 µl to 1 µl volumes
Adjustable 1-25 µl pipettes for reagent preparation
100 µl and 1 liter graduated cylinders
Tubes to prepare standard and sample dilutions
Absorbent paper
Microplate reader capable of measuring absorbance at 450nm
Log-log graph paper or computer and software for ELISA data analysis

Protocol Outline

Prepare all reagents, samples and standards as instructed in the manual.
Add 100 µl of standard or sample to each well.
Incubate 2.5 h at RT or O/N at 4°C.
Add 100 µl of prepared biotin antibody to each well.
Incubate 1 h at RT.
Add 100 µl of prepared Streptavidin solution to each well.
Incubate 45 min at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl of Stop Solution to each well.
Read at 450 nm immediately.

Need your results faster? Try RayBiotech's SpeedELISA platform for quantitative detection in just three hours.

Storage/Stability

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Documents

Datasheet (ELR-TNFa.pdf)

Datasheet (ELR-TNFa-CL.pdf)

SDS (ELR-TNFa.pdf)

SDS (ELR-TNFa-CL.pdf)

Excel (sandwich-elisa.xlsm)

Citations

Oyewo, Emmanuel Bukoye, and Musbau Adewumi Akanji. "Immune modulation potentials of aqueous extract of Andrographis paniculata leaves in male rat."?Researcher?3.1 (2011): 48-57.

Species:

Rat

Sample Type:

Serum

View Publication

Knapp, Darin J., et al. "Cytokine involvement in stress may depend on corticotrophin releasing factor to sensitize ethanol withdrawal anxiety." Brain, behavior, and immunity 25 (2011): S146-S154.

Species:

Rat

Sample Type:

Tissue lysate

View Publication

Sachdeva C., Mishra N., Sharma S. Development and characterization of enteric-coated microparticles of biochanin A for their beneficial pharmacological potential in estrogen deficient-hypertension. Drug Deliv. 2016 Jul;23(6):2044-57. doi: 10.3109/10717544.2015.1114046.

Species:

Rat

Sample Type:

Tissue Lysate (Aortic tissue lysate)

View Publication

Allabassi M. The Possible Protective Effects of Zileuton against Pulmonary Fibrosis Induced by Amiodarone in Male Rats. Journal of Natural Sciences Research. Vol.5, No.2, 2015

Species:

Rat

Sample Type:

Tissue Lysate (Lung tissue treated with Zileuton)

View Publication

Abdelkader NF., Safar MM., Salem H. Ursodeoxycholic Acid Ameliorates Apoptotic Cascade in the Rotenone Model of Parkinson's Disease: Modulation of Mitochondrial Perturbations. Mol Neurobiol. 2014 Dec 15. [Epub ahead of print]. DOI 10.1007/s12035-014-9043-8

Species:

Rat

Sample Type:

Tissue Lysate (Striatal brain tissue samples)

View Publication

Tang J., Chen Q., Guo J., et al. Minocycline Attenuates Neonatal Germinal-Matrix-Hemorrhage-Induced Neuroinflammation and Brain Edema by Activating Cannabinoid Receptor 2. Mol Neurobiol. 2015 Apr 2. epub ahead of print

Species:

Rat

Sample Type:

Tissue Lysate (Brain lysates in matrix Hemorrhage-Induced mice)

View Publication

Hedya SA, Safar MM, Bahgat AK. Cilostazol Mediated Nurr1 and Autophagy Enhancement: Neuroprotective Activity in Rat Rotenone PD Model. Mol Neurobiol. 2018 Sep;55(9):7579-7587. doi: 10.1007/s12035-018-0923-1. Epub 2018 Feb 10. PMID: 29429051.

Species:

Rat

Sample Type:

Tissue Lysate (Striatal Tissue)

View Publication

Ikutomi M., Sahara M., Nakajima T., et al. Diverse contribution of bone marrow-derived late-outgrowth endothelial progenitor cells to vascular repair under pulmonary arterial hypertension and arterial neointimal formation. J Mol Cell Cardiol. 2015 Sep;86:121-35. doi: 10.1016/j.yjmcc.2015.07.019.

Species:

Rat

Sample Type:

Conditioned Media (Endothelial progenitor cells, Rat endothelial aoritc cells, AoECS, and BM-derived mononuclear cells)

View Publication

Chen B., Zhou W., Ning M., et al. Evaluation of antitumour activity of tea carbohydrate polymers in hepatocellular carcinoma animals. Int J Biol Macromol. 2012 May 1;50(4):1103-8. doi: 10.1016/j.ijbiomac.2012.03.001.

Species:

Human

Sample Type:

Serum (Hepatocellular carcinoma rat model treated with tea carbohydrate)

View Publication

Bhardwaj, M., Padmavathi, T. K., Mani, S., Malarvizhi, R., Sali, V. K., & Vasanthi, H. R. (2020). Sulfated polysaccharide from Turbinaria ornata suppress lipopolysaccharide-induced inflammatory response in RAW 264.7 macrophages. International Journal of Biological Macromolecules. doi:10.1016/j.ijbiomac.2020.09.036

Species:

Rat

Sample Type:

Conditioned media (cell supernatant)

View Publication

View All Publications For This Product

Reviews

Write a customer review

Pretty simple kit

Rating
80%

The kit is easy enough to use. I train most people in the lab and I would say the only thing that gives it 4 stars is because the labeling of the components. With so many different letters used for one item (ie. Assay diluent A (item D), it seems to be a struggling point with new students learning the process.

Review by Decent Assay,

from UNMC,

on Sep 7, 2023
Accurate and Understandable

Rating
100%

The purchasing and shipping was no problem. We used this for a high fat rat study and it was effective at the lower limits of the detection range.

Review by Anthony,

from Pennsylvania State University ,

on Jun 9, 2023
We measured plasma levels of TNF-a. The standard curve was good, but we could not detect. Our rats were casted for 14 days and muscle atrophy was occured.

Rating
20%

14 days casting induced muscle atrophy; muscles significantly decreased muscle mass, but could not detect plasma levels of TNF-alpha

Review by Cytokines1,

from University of Texas at San Antonio,

on May 26, 2023
Great product

Rating
100%

We used the rat TNF-alpha ELISA kit to determine concentrations of TNF alpha in our rat plasma (heparin anticoagulated) and kidney extract samples. After determining optimal concentrations using two strips, ran the entire plate and everything worked perfectly. We were able to corroborated results from other experiments.

Review by Nancy,

from University of Wisconsin,

on Feb 25, 2021
Great product

Rating
80%

Very accurate and easy to follow the steps.

Review by Vcsantos,

from University of Washington,

on Jan 18, 2020
Great product

Rating
100%

Kit works very well, easy to use, excellent results.

Review by Jillian A.,

from University of Georgia,

on Jan 17, 2020
Great product

Rating
100%

Kit works really well.

Review by Karthikeyan Thirugnanam,

from Medical College of Wisconsin,

on Aug 20, 2019
Great product

Rating
100%

Excellent results; easy to use; reproducible

Review by Kay Beharry,

from SUNY Downstate Medical Center,

on Jul 30, 2019

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