Mouse TNF-alpha ELISA

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

ELM-TNFa-1
+

Antigen Information

Accession Number:
  • P06804
Gene ID:
  • 21926
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]
Target Species:
  • Mouse

Assay Format

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

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Specificity:
This ELISA kit shows no cross-reactivity with any of the cytokines tested: Mouse CD30, L CD30, T CD40, CRG-2, CTACK, CXCL16, Eotaxin , Eotaxin-2, Fas Ligand, Fractalkine, GCSF, GM-CFS, IFN- gamma, IGFBP-3, IGFBP-5, IGFBP-6, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-3 Rb, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17, KC, Leptin R, LEPTIN(OB), LIX, L-Selectin, Lymphotactin, MCP-1, MCP-5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-2, MIP-3 beta, MIP-3 alpha, PF-4, P-Selectin, RANTES, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TIMP-1, TNF RI, TNF RII, TPO, VCAM-1, VEGF.
Number of Targets Detected:
1
Species Detected:
  • Mouse
Compatible Sample Types:
  • Cell Culture Supernatants
  • Plasma
  • Serum
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. Gong J., Liu H., Wu J., et al. Maresin 1 prevents lipopolysaccharide-induced neutrophil survival and accelarates resolution of acute lung injury. Shock. 2015 Oct;44(4):371-80. doi: 10.1097/SHK.0000000000000434.
    Species: Mouse
    Sample type: BAL Fluid
  2. Choudhury S, Ghosh S., Gupta P., et al. Inflammation-induced ROS generation causes pancreatic cell death through modulation of Nrf2/NF-?B and SAPK/JNK pathway. Free Radic Res. 2015;49(11):1371-83. doi: 10.3109/10715762.2015.1075016
    Species: Mouse
    Sample type: Serum
  3. Barbhuiya S., Sengupta M. The Toxic Effects of Lead on Testicular Macrophage Immunomodualation and Sperm Cell Parameters in Mice. 9(5): XX-XX, 2015, Article no.BJMMR.17937 ISSN: 2231-0614
    Species: Mouse
    Sample type: Conditioned Media
  4. Kabel A., Omar M., Balaha M., Borg H. Effect of metformin and adriamycin on transplantable tumor model. Tissue Cell. 2015 Oct;47(5):498-505. doi: 10.1016/j.tice.2015.07.003.
    Species: Mouse
    Sample type: Tissue Lysate
  5. Mahanti S., Majhi A., Kundu K., et al. Systemic Staphylococcus aureus infection in restraint stressed mice modulates impaired immune response resulting in improved behavioral activities. J Neuroimmunol. 2015 Nov 15;288:102-13. doi: 10.1016/j.jneuroim.2015.09.003
    Species: Mouse
    Sample type: Serum
  6. Mrvova N., Skandik M., Kuniakova M., Rackova L. Modulation of BV-2 microglia functions by novel quercetin pivaloyl ester. Neurochem Int. 2015 Nov;90:246-54. doi: 10.1016/j.neuint.2015.09.005.
    Species: Mouse
    Sample type: Conditioned Media
  7. Abood A., Elshal M. VDR stimulation improves outcome of isoprenaline-induced myocardial infarction in rats via down-regulation of cardiac inos gene expression. Biomedical Research 2015; 26 (4): 755-764
    Species: Mouse
    Sample type: Serum
  8. Nandi A., Bishayi B. Host antioxidant enzymes and TLR-2 neutralization modulate intracellular survival of Staphylococcus aureus: Evidence of the effect of redox balance on host pathogen relationship during acute staphylococcal infection. Microb Pathog. 2015 Dec;89:114-27. doi: 10.1016/j.micpath.2015.09.007.
    Species: Mouse
    Sample type: Conditioned Media
  9. Bahr H., Torahi E., Mohammed E., et al. Chemopreventive effect of leflunomide against Ehrlich's solid tumor grown in mice: Effect on EGF and EGFR expression and tumor proliferation. Life Sci. 2015 Nov 15;141:193-201. doi: 10.1016/j.lfs.2015.10.003.
    Species: Mouse
    Sample type: Serum
  10. Zhang X., Zhou Y., Zhang J., et al. IL-35 inhibits acute graft-versus-host disease in a mouse model. Int Immunopharmacol. 2015 Dec;29(2):383-92. doi: 10.1016/j.intimp.2015.10.025
    Species: Mouse
    Sample type: Tissue Lysate
The kit was packaged in an organized and clear manner, and it was easy to walk through each step of the procedure. I used the mouse IL-6, VEGF, and TNF-alpha kits for plasma as well as homogenized muscle samples. Each worked well, and I would be happy to use them again.

Houston Methodist Research Institute
This kit worked very well for the analysis of mouse serum samples. It was shipped quickly, easy to use, and did exactly what it was supposed to. I highly recommend this product!

BTS Research
I used this ELISA kit for Serum and cell culture supernatant analysis. Great product!
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Usage

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

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