ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)

Brand

Cloud-Clone

short description

Enzyme-linked immunosorbent assay for Antigen Detection.
Research focus: Cytokine; Tumor immunity; Infection immunity; ;

spezie

Mus musculus (Mouse)

Cross Reaction

This assay has high sensitivity and excellent specificity for detection of Tumor Necrosis Factor Alpha (TNFa).
No significant cross-reactivity or interference between Tumor Necrosis Factor Alpha (TNFa) and analogues was observed.

detection target

Tumor Necrosis Factor Alpha

alias

DIF; TNF-A; TNFSF2; Cachectin; Tumor Necrosis Factor Ligand Superfamily Member 2

sensitivity

The minimum detectable dose of this kit is typically less than 6.1 pg/mL

detection range

15.6-1,000 pg/mL

certificate

ISO 9001:2008, ISO 13485:2003

sample type

serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids

pack size

48T, 96T, 5x96T, 10x96T

Reproducibility

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tumor Necrosis Factor Alpha (TNFa) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Tumor Necrosis Factor Alpha (TNFa) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Assay time

3h

storage and Stability

All the kits should stored according to the manual.
The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 degree upon receipt while the others should be at 4 degree.
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Principle of the Elisa

Double-antibody Sandwich
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Tumor Necrosis Factor Alpha (TNFa). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Tumor Necrosis Factor Alpha (TNFa). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Tumor Necrosis Factor Alpha (TNFa), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Tumor Necrosis Factor Alpha (TNFa) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Kit Content

Pre-coated, ready to use 96-well strip plate: 1
Plate sealer for 96 wells: 4
Standard: 2
Standard Diluent: 1x20mL
Detection Reagent A: 1x120µ L
Assay Diluent A: 1x12mL
Detection Reagent B: 1x120µ L
Assay Diluent B: 1x12mL
TMB Substrate: 1x9mL
Stop Solution: 1x6mL
Wash Buffer (30 x concentrate): 1x20mL
Instruction manual: 1

Assay procrdure summery

1. Prepare all reagents, samples and standards;
2. Add 100µ L standard or sample to each well. Incubate 2 hours at 37° C;
3. Aspirate and add 100µ L prepared Detection Reagent A. Incubate 1 hour at 37° C;
4. Aspirate and wash 3 times;
5. Add 100µ L prepared Detection Reagent B. Incubate 30 minutes at 37° C;
6. Aspirate and wash 5 times;
7. Add 90µ L Substrate Solution. Incubate 10-20 minutes at 37° C;
8. Add 50µ L Stop Solution. Read at 450nm immediately.

Reference

Deficiency of Rac1 Blocks NADPH Oxidase Activation, Inhibits Endoplasmic Reticulum Stress, and Reduces Myocardial Remodeling in a Mouse Model of Type 1 Diabetes.; Protective Effect of Nicotine on Lipopolysaccharide-Induced Acute Lung Injury in Mice; Mesangial medium with IgA1 from IgA nephropathy inhibits nephrin expression in mouse podocytes; Combined Effects of Hematopoietic Progenitor Cell Mobilization from Bone Marrow by Granulocyte Colony Stimulating Factor and AMD3100 and Chemotaxis into the Brain Using Stromal Cell-Derived Factor-1? in an Alzheimer' s Disease Mouse Model; Egr-1 decreases adipocyte insulin sensitivity by tilting PI3K/Akt and MAPK signal balance in mice; Ixora coccinea down Regulate the Inflammation-Associated Prostaglandins (PGE-2), Nitric Oxide (NO) and Cytokines Production in LPS-Stimulated RAW 264.7 Macrophage; Tumor necrosis factor ? (TNF-?) receptor-I is required for TNF-?-mediated fulminant virus hepatitis caused by murine hepatitis virus strain-3 infection.; Protective effect of marine mangrove Rhizophora apiculata on acetic acid induced experimental colitis by regulating anti-oxidant enzymes, inflammatory mediators and nuclear factor-kappa B subunits; Anti-inflammatory Effect of Euphorbia hirta Leaf Extract in Rats and Modulation of Inflammation-Associated Prostaglandins (PGE-2) and Nitric Oxide (NO) Expression in RAW264.7 Macrophage; Biophysico-chemical interfacial attributes of Fe3O4 decorated MWCNT nanohybrid/bio-based hyperbranched polyurethane nanocomposite: an antibacterial wound healing material with controlled drug release potential; Ginsenosides Rg1 from Panax ginseng: A Potential Therapy for Acute Liver Failure Patients?; Tumor necrosis factor ? (TNF-?) receptor-I is required for TNF-?-mediated fulminant virus hepatitis caused by murine hepatitis virus strain-3 infection; Valproic acid protects septic mice from renal injury by reducing the inflammatory response; Involvement of nitric oxide with activation of Toll-like receptor 4 signaling in mice with dextran sodium sulfate-induced colitis; Differential Pro-Inflammatory Responses of Astrocytes and Microglia Involve STAT3 Activation in Response to 1800 MHz Radiofrequency Fields; Dietary squid ink polysaccharide could enhance SIgA secretion in chemotherapeutic mice; Protective effects of astaxanthin against ischemia/reperfusion induced renal injury in mice; Exogenous carcinoembryonic antigen-related cell adhesion molecule 1 suppresses 2,4,6-trinitrobenzene sulfonic acid-induced ulcerative colitis in mice; Matrix Metalloproteinase 9 Secreted by Hypoxia Cardiac Fibroblasts Triggers Cardiac Stem Cell Migration In Vitro; Irisin improves endothelial function in obese mice through the AMPK-eNOS pathway; Protein Inhibitor of Activated STAT 1 (PIAS1) Protects Against Obesity-Induced Insulin Resistance by Inhibiting Inflammation Cascade in Adipose Tissue; Epigallocatechin-3-Gallate Attenuates Unilateral Ureteral Obstruction-Induced Renal Interstitial Fibrosis in Mice; MicroRNA-155 Promotes Atherosclerosis Inflammation via Targeting SOCS1; Trans?fatty acid promotes thrombus formation in mice by aggravating antithrombogenic endothelial functions via Toll?like receptors; Trypanosoma cruzi infection and benznidazole therapy independently stimulate oxidative status and structural pathological remodeling of the liver tissue in mice; Impaired adipogenesis in adipose tissue associated with hepatic lipid deposition induced by chronic inflammation in mice with chew diet; Fibronectin type III domain containing 5 expression in skeletal muscle in chronic heart failureﾗrelevance of inflammatory cytokines; Hypoxia-induced secretion of IL-15 from adipose-derived mesenchymal stem cell promotes growth and cancer stem cell properties of Burkitt lymphoma; Modulation of oxidative and inflammatory cardiac response by nonselective 1?and 4?cyclooxygenase inhibitor and benznidazole in mice; Construction of recombinant Mip-FlaA dominant epitope vaccine against Legionella pneumophila and evaluation of the immunogenicity and protective immunity; Protective role of n6/n3 PUFA supplementation with varying DHA/EPA ratios against atherosclerosis in mice; Effects of Hyriopsis cumingii Polysaccharides on Mice Immunologic Receptor, Transcription Factor, and Cytokine; Lipopolysaccharide Cross-Tolerance Delays Platelet-Activating Factor-Induced Sudden Death in Swiss Albino Mice: Involvement of Cyclooxygenase in Cross-Tolerance; Effects of Dectin-1 on the mast cells in allergic conjunctivitis and its underlying mechanism;