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. Bishayi B., et al. Effect of exogenous MCP-1 on TLR-2 neutralized murine macrophagesand possible mechanisms of CCR-2/TLR-2 and MCP-1 signallingduring Staphylococcus aureus infection. Immunobiology Ahead of Print 2014, DOI:10.1016/j.imbio.2014.10.013
    Species: Mouse
    Sample type: Conditioned Media
  2. Dey S., Majhi A., Mahanti S., et al. In Vitro Anti-inflammatory and Immunomodulatory Effects of Ciprofloxacin or Azithromycin in Staphylococcus aureus-Stimulated Murine Macrophages are Beneficial in the Presence of Cytochalasin D. Inflammation, epub ahead of print Nov 2014. DOI: 10.1007/s10753-014-0070-4
    Species: Mouse
    Sample type: Conditioned Media
  3. Joshi N., Shirsath N., Sing A., et al. Endogenous lung surfactant inspired pH responsive nanovesicle aerosols: Pulmonary compatible and site-specific drug delivery in lung metastases. Scientific Reports 4, Article number:7085, doi:10.1038/srep07085
    Species: Mouse
    Sample type: Conditioned Media
  4. Joshi, N., Shirsath, N., Singh, A., Joshi, K.S. & Banerjee, R. Endogenous lung surfactant inspired pH responsive nanovesicle aerosols: Pulmonary compatible and site-specific drug delivery in lung metastases. Sci. Rep. 4, 7085; DOI:10.1038/srep07085 (2014).
    Species: Mouse
    Sample type: Conditioned Media
  5. Majhi A., Ashikary R., Bhattacharyya A., Mahanti S., Bishayi B. LEVOFLOXACIN AND CEFTRIAXONE IN COMBINATION ATTENUATES LUNG INFLAMMATION IN A MOUSE MODEL OF BACTEREMIC PNEUMONIA BY MULTI-DRUG RESISTANT Streptococcus pneumoniae VIA INHIBITION OF CYTOLYTIC ACTIVITIES OF PNEUMOLYSIN AND AUTOLYSIN. Antimicrob. Agents Chemother. Accepted ahead of Print, June 23 2014. doi:10.1128/AAC.03245-14
    Species: Mouse
    Sample type: Serum
  6. Meng A., et al.Role of p38 MAPK and STAT3 in lipopolysaccharide?stimulated mouse alveolar macrophages. EXPERIMENTAL AND THERAPEUTIC MEDICINE 8: 1772-1776, 2014. DOI: 10.3892/etm.2014.2023
    Species: Mouse
    Sample type: Conditioned Media
  7. Morganti J., Jopson T., Liu S., Gupta N., Rosi S. Cranial Irradiation Alters the Brain?s Microenvironment and Permits CCR2+ Macrophage Infiltration. PLOS One Published: April 02, 2014 DOI: 10.1371/journal.pone.0093650 [Epub Ahead of Print]
    Species: Mouse
    Sample type: Tissue Lysate
  8. Moulari, B., Beduneau, A. et al. Lectin-decorated nanoparticles enhance binding to the inflamed tissue in experimental colitis. Journal of Controlled Release 2014. Epub Ahead of Print. DOI: 10.1016/j.jconrel.2014.05.046
    Species: Mouse
    Sample type: Colon Fluid
  9. Shi Q., Cao J., Fang L., Zhao., Liu Z., Ran., et al. Geniposide suppresses LPS-induced nitric oxide, PGE< sub> 2 and inflammatory cytokine by downregulating NF-?B, MAPK and AP-1 signaling pathways in macrophages. Int Immunopharmacol 2014 Apr 13;20(2):298-306. Epub 2014 Apr 13.
    Species: Mouse
    Sample type: Conditioned Media
  10. Singh JC., Kakalij RM., Kshirsagar RP., et al. Cognitive effects of vanillic acid against streptozotocin-induced neurodegeneration in mice. Pharm Biol. 2014 Dec 4:1-7
    Species: Mouse
    Sample type: Serum
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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