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

Oryctolagus cuniculus (Rabbit)

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 1.17 pg/mL

detection range

3.12-200 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

P04924

Reference

Enhanced tumor growth after portal vein embolization in a rabbit tumor model; Comparison of Acute Effect of Systemic Versus Intravitreal Infliximab Treatment in an Experimental Model of Endotoxin-Induced Uveitis; Repeated open endotracheal suctioning causes gradual desaturation but does not exacerbate lung injury compared to closed endotracheal suctioning in a rabbit model of ARDS.; Budesonide added to modified porcine surfactant Curosurf may additionally improve the lung functions in meconium aspiration syndrome.; Immunosuppression abrogates resistance of young rabbits to Rabbit Haemorrhagic Disease (RHD); Nonalcoholic steatohepatitis as a novel player in metabolic syndrome-induced erectile dysfunction: An experimental study in the rabbit; Hyperlipidemia causes changes in inflammatory responses to periodontal pathogen challenge: Implications in acute and chronic infections; Bisphenol A Exposure Enhances Atherosclerosis in WHHL Rabbits; Systemic and Flap Inflammatory Response Associates with Thrombosis in Flap Venous Crisis; Panax notoginseng saponins promote wound repair of anterior cruciate ligament through phosphorylation of PI3K, AKT and ERK; ??????????????????????????; Effects of Intravenous Injection of; Alterations of the thioredoxin system during subarachnoid hemorrhage-induced cerebral vasospasm; Effects of morphine on the expression of cytokines and inflammatory mediators in a rabbit model of endotoxin-induced experimental uveitis; Differential Interactive Effects of Cartilage Traumatization and Blood Exposure In Vitro and In Vivo; ???????? ???????????? ??????? ? ?????? ????????????? ?????? ??????? ?????????? ????????????? ??????????? ? ????????????; Hyperinflation deteriorates arterial oxygenation and lung injury in a rabbit model of ARDS with repeated open endotracheal suctioning; The C-terminal Domain Supports a Novel Function for CETPI as a New Plasma Lipopolysaccharide-Binding Protein; Telmisartan ameliorates oxidative stress and subarachnoid haemorrhage-induced cerebral vasospasm;