Mouse TNF-alpha ELISA

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ELM-TNFa-1

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

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Antigen Information

Accession Number:
  • P06804
Gene ID:
  • 21926
Gene Symbol:
  • 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:
  • 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. Chandan R., Banerjee R. Pro-apoptotic liposomes-nanobubble conjugate synergistic with paclitaxel: a platform for ultrasound responsive image-guided drug delivery. Sci Rep. 2018 Feb 8;8(1):2624. doi: 10.1038/s41598-018-21084-8.
    Species: Mouse
    Sample type: Conditioned Media
  2. Wang J., Sun Z., Gou W., et al. ?-1 Antitrypsin Enhances Islet Engraftment by Suppression of Instant Blood-Mediated Inflammatory Reaction. Diabetes. 2017 Apr;66(4):970-980. doi: 10.2337/db16-1036.
    Species: Mouse
    Sample type: Serum
  3. Rasheed N., Ahmed L., Abdallah D., El-Sayed B. Nephro-toxic effects of intraperitoneally injected EGCG in diabetic mice: involvement of oxidative stress, inflammation and apoptosis. Sci Rep. 2017; 7: 40617. doi: 10.1038/srep40617
    Species: Mouse
    Sample type: Serum
  4. Zhang S., Wang Y., Li D., et al. Necrostatin-1 Attenuates Inflammatory Response and Improves Cognitive Function in Chronic Ischemic Stroke Mice. Medicines 2016, 3(3), 16; doi:10.3390/medicines3030016
    Species: Mouse
    Sample type: Tissue Lysate
  5. Yu J., Yuan X., Liu Y., et al. Delayed Administration of WP1066, an STAT3 Inhibitor, Ameliorates Radiation-Induced Lung Injury in Mice. Lung (2016) 194: 67. doi:10.1007/s00408-015-9821-8
    Species: Mouse
    Sample type: Serum
  6. Di Cristo L., Movia D., Bianchi M., et al. Proinflammatory Effects of Pyrogenic and Precipitated Amorphous Silica Nanoparticles in Innate Immunity Cells. Toxicol Sci. 2016 Mar;150(1):40-53. doi: 10.1093/toxsci/kfv258
    Species: Mouse
    Sample type: Conditioned Media
  7. Nandi A., Bishayi B. Murine macrophage response from peritoneal cavity requires signals mediated by chemokine receptor CCR-2 during Staphylococcus aureus infection. Immunol Res. 2016 Feb;64(1):213-32. doi: 10.1007/s12026-015-8739-9.
    Species: Mouse
    Sample type: Conditioned Media
  8. Dey P., Ray S., Chaudhuri T. Immunomodulatory activities and phytochemical characterisation of the methanolic extract of Dioscorea alata aerial tuber. Journal of Functional Foods . Volume 23, May 2016, Pages 315-328. https://doi.org/10.1016/j.jff.2016.02.044
    Species: Mouse
    Sample type: Conditioned Media
  9. Bianchi M., Allegri M., Costa A., et al. Titanium dioxide nanoparticles enhance macrophage activation by LPS through a TLR4-dependent intracellular pathway. Toxicol. Res., 2015,4, 385-398 DOI: 10.1039/C4TX00193A
    Species: Mouse
    Sample type: Conditioned Media
  10. Dey P., Chaudhuri TK. Immunomodulatory activity of Nerium indicum through inhibition of nitric oxide and cyclooxygenase activity and modulation of TH1/T H2 cytokine balance in murine splenic lymphocytes. Cytotechnology. 2015 Feb 14. [Epub ahead of print]
    Species: Mouse
    Sample type: Conditioned Media
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.