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

Homo sapiens (Human)

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

15.6-1000 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.

Uniport ID

P01375

Reference

Analysis of genetic markers for cardiovascular disorders in a portuguese population with familial hypercholesterolemia; Soluble BAFF-R produced by decidual stromal cells plays an inhibitory role in monocytes and macrophages ; Biomarkers of Fibroproliferative Healing in Fibrosing Idiopathic Interstitial Pneumonias; A Combinatorial Relative Mass Value Evaluation of Endogenous Bioactive Proteins in Three-Dimensional Cultured Nucleus Pulposus Cells of Herniated Intervertebral Discs: Identification of Potential Target Proteins for Gene Therapeutic Approaches; Immunogenic alteration in laparoscopic common bile duct exploration.; Effects of Photodynamic Therapy on the Clinical and Gingival Crevicular Fluid Inflammatory Biomarkers in Chronic Periodontitis: A Split-Mouth Randomized Clinical Trial; PAR?2, IL?4R, TGF?? and TNF?? in bronchoalveolar lavage distinguishes extrinsic allergic alveolitis from sarcoidosis; Increased serum levels of lipocalin-1 and-2 in patients with stable chronic obstructive pulmonary disease; Metformin Inhibits the IL-6-Induced Epithelial-Mesenchymal Transition and Lung Adenocarcinoma Growth and Metastasis; GYY4137, a hydrogen sulfide?releasing molecule, inhibits the inflammatory response by suppressing the activation of nuclear factor?kappa B and mitogen?activated protein kinases in Coxsackie virus B3?infected rat cardiomyocytes.; Systemic and Flap Inflammatory Response Associates with Thrombosis in Flap Venous Crisis; Sulfiredoxin-1 protects primary cultured astrocytes from ischemia-induced damage; Estrogen-deficient osteoporosis enhances the recruitment and activity of osteoclasts by breast cancer cells; MicroRNA-155 Promotes Atherosclerosis Inflammation via Targeting SOCS1; MicroRNA-155 Promotes Atherosclerosis Inflammation via Targeting SOCS1; The immunomodulating effect of seminal plasma on T cells; Effects of different ventilation strategies on exhaled nitric oxide in geriatric abdominal surgery; Low Shear Stress Induced HMGB1 Translocation and Release via PECAM-1/PARP-2 Pathway to Induce Inflammation Response; ??????????????????????????????????; Cellular uptake of exogenous calcineurin B is dependent on TLR4/MD2/CD14 complexes, and CnB is an endogenous ligand of TLR4; Role of reactive oxygen species-mediated MAPK and NF-?B activation inﾠpolygonatum cyrtonemalectin-induced apoptosis and autophagy in human lung adenocarcinoma A549 cells; Neutrophil gene dynamics and plasma cytokine levels in dairy cattle during peri-implantation period; Human mesenchymal stem cells attenuate experimental bronchopulmonary dysplasia induced by perinatal inflammation and hyperoxia; Influence of diet on leukocyte telomere length, markers of inflammation and oxidative stress in individuals with varied glucose tolerance: a Chinese population study; Investigation of the anti-glioma activity of Oviductus ranae protein hydrolysate; Plasma Periostin Levels Are Increased in Chinese Subjects with Obesity and Type 2 Diabetes and Are Positively Correlated with Glucose and Lipid Parameters;