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Human TNF-alpha ELISA

RayBio® Human TNF alpha ELISA Kit for cell culture supernatants, plasma, serum, and lysate samples.

$402.00  
ELH-TNFa-1
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Antigen Information

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]
Target Species:
  • Human

Assay Format

Detection Range:
30 pg/ml - 6000 pg/ml
Recommended Dilution (serum & plasma):
2 fold
Sensitivity:
30 pg/ml

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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
Number of Targets Detected:
1
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Cell Lysates
  • Plasma
  • Serum
  • Tissue Lysates
Design Principle:
  • Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
  • Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

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

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.
  1. Siqi Feng, Ya Su, Li Luo, Jing Fengchuan, Qijian Yi, Serum levels of C1q/tumor necrosis factor related protein-1 in children with Kawasaki disease, Pediatric Research accepted article preview 23 January 2018; doi: 10.1038/pr.2018.16.
    Species: Human
    Sample type: Serum
  2. Sambataro M, Sambado L., Trevisiol E., et al. Proinsulin-expressing dendritic cells in type 2 neuropathic diabetic patients with and without foot lesions. FASEB J. 2018 Feb 12:fj201701279RR. doi: 10.1096/fj.201701279RR
    Species: Human
    Sample type: Plasma
  3. Habashy N., Abu Serie M., Attia W., Abdelgaleil S. Chemical characterization, antioxidant and anti-inflammatory properties of Greek Thymus vulgaris extracts and their possible synergism with Egyptian Chlorella vulgaris. Journal of Functional Foods , Volume 40, January 2018, Pages 317-328, doi.org/10.1016/j.jff.2017.11.022
    Species: Human
    Sample type: Conditioned Media
  4. Yamini, P., R. S. Ray, and Kanwaljit Chopra. "Vitamin D 3 attenuates cognitive deficits and neuroinflammatory responses in ICV-STZ induced sporadic Alzheimer?s disease."?Inflammopharmacology?26.1 (2018): 39-55.
    Species: Human
    Sample type: Tissue
  5. Sukoyan G., Ionov D., Galenko-Iaroshevsky P., et al. Severity of Symptoms of Systemic Inflammatory Response Syndrome and Congestive Heart Failure Caused by Chronic Aortic Stenosis and Its Pharmacological Correction. World Journal of Cardiovascular Diseases, 2017, 7, 465-476
    Species: Rabbit
    Sample type: Tissue Lysate
  6. 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: Cell culture supernatants
  7. Hussein N., Sharara G. Correlation between serum leptin, cytokines, cartilage degradation and functional impact in obese knee osteoarthritis patients. The Egyptian Rheumatologist. Volume 38, Issue 2, April 2016, Pages 117-122
    Species: Human
    Sample type: Serum
  8. Liu Z., Sun C., Tao R., et al. Pyrroloquinoline Quinone Decelerates Rheumatoid Arthritis Progression by Inhibiting Inflammatory Responses and Joint Destruction via Modulating NF-?B and MAPK Pathways. Inflammation. 2016 Feb;39(1):248-56. doi: 10.1007/s10753-015-0245-7.
    Species: Human
    Sample type: Conditioned Media
  9. Zhang L., Wang X., Miao Y., et al. Magnetic ferroferric oxide nanoparticles induce vascular endothelial cell dysfunction and inflammation by disturbing autophagy. J Hazard Mater. 2016 Mar 5;304:186-95. doi: 10.1016/j.jhazmat.2015.10.041
    Species: Human
    Sample type: Conditioned Media
  10. Yu Y., Li X., Qu L., et al. DXXK exerts anti-inflammatory effects by inhibiting the lipopolysaccharide-induced NF-?B/COX-2 signalling pathway and the expression of inflammatory mediators. J Ethnopharmacol. 2016 Feb 3;178:199-208. doi: 10.1016/j.jep.2015.11.016.
    Species: Human
    Sample type: Conditioned Media
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Great kit. Easy to follow protocol. Getting great results.
Patricia
Florida A&M University 02/12/2020, 02:05 PM
It's so easy to use. I'm happy with it and recommend it to everyone.
George
Columbia University 01/21/2020, 10:37 AM
Simple protocol and reproducible results. Would recommend.
Naureen Fatima
University of Houston 05/04/2019, 02:51 PM
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.
DHARA SHAH
University of Illinois at Chicago 06/29/2018, 09:22 AM
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# Manual (Supernatants / Plasma / Serum)

# Manual (Lysates)

# SDS

Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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