Size	1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Species	Human
Accession Number	P01375
Gene Id	7124
Gene Symbols	TNF\|TNFA\|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: Human ANG, CD23, Eotaxin, GCSF, GM-CSF, GRO-alpha, GRO-beta, GRO-gamma, I-309, IFN-gamma, IL-1 alpha, IL-1 beta, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12 (p70), IL-15, IL-16, IP-10, MCP-1, MCP-2, MCP-3, MCP-4, MCSF, MIG, MIP-1 alpha, MIP-1 beta, NAP-2, PDGF, PF-4
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
Research Area	Antibody Drug Development, Protein Therapeutics, Biomarker Detection
Sensitivity	30 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	30 pg/ml - 6000 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

Standard curve graphs showing the results from the human tnf alpha elisa kit

Dilution series diagram with 8 vials

Spiking & Recovery Results

Sample Type	Average % Recovery	Range (%)
Serum	94.53	82-103
Plasma	91.92	80-102
Cell culture media	96.43	84-104

Linearity Results

		Serum	Plasma	Cell Culture Media
1:2	Average % of expected Range (%)	94 83-102	93 82-103	92 81-102
1:4	Average % of expected Range (%)	94 83-104	92 82-103	99 85-107

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

Manual (ELH-TNFa.pdf)

Manual (ELH-TNFa-CL.pdf)

SDS (ELH-TNFa.pdf)

Excel (sandwich-elisa.xlsm)

Citations

Chen, Xia, et al. "LTM chloride inhibits LPS mediated expressions of pro-inflammatory cytokines and adhesion molecules in HUVECs: involvement of NF-kB-Jmjd3 signaling." Cellular Physiology and Biochemistry 42.5 (2017): 1713-1724.

Species:

Human

Sample Type:

Cell culture supernatants

View Publication

Mause S., Ritzel E., Liehn E., et al. Platelet Microparticles Enhance the Vasoregenerative Potential of Angiogenic Early Outgrowth Cells After Vascular Injury. Circulation . 2010 Aug 3;122(5):495-506. doi: 10.1161/CIRCULATIONAHA.109.909473.

Species:

Human

Sample Type:

Conditioned Media (HUVEC cells in vascular injury model)

View Publication

Gao Q., Camous X., Lu Y., et al. Novel inflammatory markers associated with cognitive performance: Singapore Longitudinal Ageing Studies. Neurobiol Aging. 2016 Mar;39:140-6. doi: 10.1016/j.neurobiolaging.2015.12.002.

Species:

Human

Sample Type:

Plasma (Cognitive Performance)

View Publication

Fermoselle C., Rabinovich R., Ausin P., et al. Does oxidative stress modulate limb muscle atrophy in severe COPD patients? Eur Respir J. 2012 Oct;40(4):851-62.

Species:

Human

Sample Type:

Tissue Lysate (Quadricep Biopsies from COPD patients)

View Publication

Jiang H., Ling Z., Zhang Y., et al. Altered fecal microbiota composition in patients with major depressive disorder. Brain Behav Immun. 2015 Aug;48:186-94. doi: 10.1016/j.bbi.2015.03.016

Species:

Human

Sample Type:

Serum (Major Depressive Disorder Patients)

View Publication

Qi C., Wang X., Shen Z., et al. Anti-mitotic chemotherapeutics promote apoptosis through TL1A-activated death receptor 3 in cancer cells. Cell Research volume 28, pages 544-555 (2018)

Species:

Human

Sample Type:

Conditioned Media (Adenocarcinoma Cell lines)

View Publication

Chen, Xia, et al. "Lanthanum chloride inhibits LPS mediated expressions of pro-inflammatory cytokines and adhesion molecules in HUVECs: involvement of NF-?B-Jmjd3 signaling." Cellular Physiology and Biochemistry 42.5 (2017): 1713-1724.

Species:

Human

Sample Type:

Conditioned Media

View Publication

Perez-Garcia JL., et al. Identification of TNF-a and MMP-9 as potential baseline predictive serum markers of sunitinib activity in patients with renal cell carcinoma using a human cytokine array. British Journal of Cancer (2009) 101, 1876 - 1883

Species:

Human

Sample Type:

Serum

View Publication

Liu Y., Liao J., Zhao M., et al. Increased expression of TLR2 in CD4(+) T cells from SLE patients enhances immune reactivity and promotes IL-17 expression through histone modifications. Eur J Immunol. 2015 Sep;45(9):2683-93. doi: 10.1002/eji.201445219.

Species:

Human

Sample Type:

Conditioned Media (CD4 T cells with a TLR2 gene expression)

View Publication

Zhenjian He, Jiahui Chen, Xun Zhu, Shu An, Xinhuai Dong, Jianchen Yu, Shihao Zhang, Yun Wu, Ge Li, Yu Zhang, Jueheng Wu, Mengfeng Li, NLRP3 Inflammasome Activation Mediates Zika Virus-Associated Inflammation, The Journal of Infectious Diseases, Volume 217, Issue 12, 15 June 2018, Pages 1942-1951, https://doi.org/10.1093/infdis/jiy131

Species:

Human

Sample Type:

Conditioned media (cell culture media)

View Publication

View All Publications For This Product

Reviews

Write a customer review

Great Product

Rating
100%

Easy to use with simple instructions to follow. A staple kit used in our research for years. Would recommend

Review by Eesha,

from AyuVis Research Inc,

on Jan 5, 2024
Great product

Rating
100%

Used it for cell culture media from prostate epithelial cells and fibroblasts. Very easy to use kit and protocol. Assay worked well.

Review by Brent,

from University of Arizona,

on Nov 16, 2023
Great product

Rating
100%

Excellent performance for the assessment of THP-1 polarization towards pro-inflammatory macrophages.

Review by Isis,

from University of Puerto Rico at Maya,

on Jul 14, 2022
Great product

Rating
100%

Excellent product, I used it for a THP inhibitory assay with great accuracy and reproducibility.

Review by Said,

from University of Puerto Rico,

on Jul 13, 2022
Great product

Rating
100%

Straightforward protocol!

Review by Evan,

from BCM,

on Jun 24, 2022
Great product

Rating
100%

Great standard curve! I used on supernatants and no needed to dilute my samples (PASMCs medium).Highly recommended

Review by Matteo Massaro,

from HMRI,

on Mar 2, 2022
Great product

Rating
100%

This ELISA kit was used for quantification of TNF-alpha in the supernatants of SEB-stimulated peripheral blood mononuclear cells in combination with conventional chemotherapeutics. Overall, the assay worked well, and the output was clear.

Review by MARINA K,

from KYNAN PHARMA,

on Jul 17, 2021
Great product

Rating
100%

Worked great with THP-1 cells stimulated with LPS. Sensitive and easy to use.

Review by Scott,

from Duke University,

on Jul 13, 2020
Great product

Rating
100%

Great kit. Easy to follow protocol. Getting great results.

Review by Patricia,

from Florida A&M University,

on Feb 11, 2020
Great product

Rating
100%

It's so easy to use. I'm happy with it and recommend it to everyone.

Review by George,

from Columbia University,

on Jan 20, 2020
Great product

Rating
100%

Simple protocol and reproducible results. Would recommend.

Review by Naureen Fatima,

from University of Houston,

on May 3, 2019
Great product

Rating
100%

I used it for human corneal epithelial cell culture supernatant under induced conditions. It is very sensitive and easy to use. I also want to comment on the fast delivery of the product.

Review by DHARA SHAH,

from University of Illinois at Chicago,

on Jun 28, 2018

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