Mouse TNF-alpha ELISA

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

$402.00  
ELM-TNFa-1
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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. Shoulders H., Farner K., Singla D. Macrophage depletionbyclondronate attenuates bonemorphogeneticprotein-7 induced M2macrophagedifferentiationand improved systolicbloodvelocityin atherosclerosis. Translational ResearchVolume 203, January 2019, Pages 1-14, in press
    Species: Serum
    Sample type: Serum
  2. 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
  3. Dey S., Bishayi B. Killing of S. aureus in murine peritoneal macrophages by Ascorbic acid along with antibiotics Chloramphenicol or Ofloxacin: Correlation with inflammation. Microb Pathog. 2018 Feb;115:239-250. doi: 10.1016/j.micpath.2017.12.048
    Species: Mouse
    Sample type: Conditioned Media
  4. 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
  5. 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
  6. Jia Y., Li Z., Cai W., et al. SIRT1 regulates inflammation response of macrophages in sepsis mediated by long noncoding RNA. Biochim Biophys Acta Mol Basis Dis. 2018 Mar;1864(3):784-792. doi: 10.1016/j.bbadis.2017.12.029.
    Species: Mouse
    Sample type: Conditioned Media
  7. Nandi, Ajeya, and Biswadev Bishayi. "A novel CCR-2/TLR-2 triggered signaling in murine peritoneal macrophages intensifies bacterial (Staphylococcus aureus) killing by reactive oxygen species through TNF-R1."?Immunology letters?190 (2017): 93-107.
    Species: Mouse
    Sample type: Cell lysates
  8. 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
  9. 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
  10. 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
This kit gave me great results with LPS-inflammed RAW264.7 cells supernatants. The instructions were clear and detailed and I like that you can use just half a plate and keep the other strips for another time. I would strongly recommend.

University of Arkansas
I used the kit for cell culture supernatant analysis. The dilution times need yourself to investigate ahead and then do lots of experiments. Good products.

ALDERSON BROADDUS UNIVERSITY
Mouse spleen lysates and with denervated muscle tissue was a sample for use of the kit. There's very little reaction with lysates but it exists. From my experience, this kit works well for cell culture supernatants.

NanoBST
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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