Catalog No: OKCD04203
Size:96WELLS
Price: $450.00
SKU
OKCD04203
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Datasheets/ManualsPrintable datasheet for TNFA ELISA Kit (Rat) (OKCD04203)
Product Info
Predicted Species ReactivityHomo sapiens|Mouse|Rat
ApplicationEnzyme-linked Immunosorbent assay-Sandwich
ELISA Kit Detection MethodColorimetric
ELISA Kit Duration3h
ELISA Kit Linearity
Sample1:21:41:81:16
Serum (n=5)81-88%92-99%92-101%79-97%
EDTA Plasma (n=5)95-104%93-101%86-94%93-105%
Heparin Plasma (n=5)85-97%88-101%78-91%97-105%
ELISA Kit PrincipleThe 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.
ELISA Kit Range15.6-1,000pg/mL
ELISA Kit Recovery
MatrixRecovery range (%)Average (%)
Serum (n=5)98-105102
EDTA Plasma (n=5)88-9691
Heparin Plasma (n=5)91-10394
ELISA Kit ReproducibilityIntra-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%
ELISA Kit Component
ComponentAmount
Tnfa Microplate96 Wells (12 x 8 Well strips)
Tnfa Lyophilized Standard2
100X Tnfa HRP-Detector Anitbody1 x 120 uL
100X Avidin-HRP Conjugate1 x 120 uL
Standard Diluent1 x 20 mL
Detector Antibody Diluent1 x 12 mL
Conjugate Diluent1 x 12 mL
30X Wash Buffer1 x 20 mL
100X Luminol Substrate1 x 2 mL
Substrate Diluent1 x 20 mL
Reconstitution and Storage2°C to 8°C|-20°C
Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Sensitivity6.1 pg/mL
Assay InfoAssay Methodology: Quantitative Sandwich ELISA
Gene SymbolTNF
Gene Full Nametumor necrosis factor
Alias SymbolsAPC1 protein;Cachectin;DIF;TNF, macrophage-derived;TNF, monocyte-derived;TNF-a;TNFA;TNF-alpha;TNFSF2;TNLG1F;tumor necrosis factor;tumor necrosis factor ligand 1F;tumor necrosis factor ligand superfamily member 2;tumor necrosis factor-alpha.
NCBI Gene Id7124
Protein NameTumor necrosis factor
Description of TargetCytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. Impairs regulatory T-cells (Treg) function in individuals with rheumatoid arthritis via FOXP3 dephosphorylation. Upregulates the expression of protein phosphatase 1 (PP1), which dephosphorylates the key 'Ser-418' residue of FOXP3, thereby inactivating FOXP3 and rendering Treg cells functionally defective (PubMed:23396208). Key mediator of cell death in the anticancer action of BCG-stimulated neutrophils in combination with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line (PubMed:22517918).
Uniprot IDP01375
Protein Accession #NP_038721.1
Nucleotide Accession #NM_013693.3
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