TNFI ELISA Kit (Mouse) (OKBB00263)
|Predicted Species Reactivity||Mouse|
|Kit Detection Method||Colorimetric, OD450 nm|
|Kit Duration||~ 3 Hours|
|Kit Principle||Aviva Systems Biology TNF ELISA Kit (Mouse) (OKBB00263) is based on standard sandwich enzyme-linked immune-sorbent assay technology. A rat monoclonal antibody specific for TNF has been pre-coated onto 96-wellplate (12 x 8 Well Strips). Standards (E.coli,L80-L235) and test samples are added to the wells, incubated and removed. A biotinylated detector antibody specific for TNF is added, incubated and followed by washing. Avidin-Biotin-Peroxidase Complex is then added, incubated and unbound conjugate is washed away. An enzymatic reaction is visualized through the addition of TMB substrate which is catalyzed by HRP to produce a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration read by absorbance at 450 nm and is quantitatively proportional to the amount of sample Mouse TNF captured in well.|
|Kit Range||15.6 pg/mL-1,000 pg/mL (cell culture supernates)
7.8 pg/mL-500 pg/mL (mouse serum, plasma)
|Additional Information||Range: 15.6pg/ml-1000pg/ml (cell culture supernates)
7.8pg/ml-500pg/ml (mouse serum, plasma)
|::||Principle: Aviva’s mouse TNFα ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Mouse TNFα specific-specific monoclonal antibodies were precoated onto 96-well plates. The mouse specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse TNFα amount of sample captured in plate.|
|::||Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
experiment using standards and a small number of samples is recommended.
2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
4. Duplicate well assay is recommended for both standard and sample testing.
5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
6. Don’t reuse tips and tubes to avoid cross contamination.
7. To avoid to use the reagents from different batches together.
8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
Background: Tumor necrosis factor-alpha (TNF-alpha, or TNF) is secreted by macrophages in response to inflammation, infection and cancer.Mouse Tumor Necrosis Factor (TNF) and Lymphotoxin (TNF-beta) are cytotoxic proteins which have similar biological activities and share 30% amino acid homology.TNF-alpha is produced by monocytes, which can stimulate endothelial cells to produce the multilineage growth factor granulocyte-macrophage colony-stimulating factor and extend the role of this immunoregulatory protein to the regulation of hematopoiesis in vitro. TNF is a soluble protein that causes damage to tumor cells but has no effect on normal cells. Mouse TNF has been purified to apparent homogeneity as a 17.3-kilodalton protein from HL-60 leukemia cells and has showed cytotoxic and cytostatic activities against various Mouse tumor cell lines. The Mouse TNF cDNA is 1585 base pairs in length and encodes a protein of 233 amino acids. The mature protein begins at residue 77, leaving a long leader sequence of 76 amino acids. TNF-alpha has been mapped to Mouse chromosome 6.
|Reconstitution and Storage||Store as indicated in product manual.|
|Sample Type||cell culture supernates, serum and plasma (heparin, EDTA)|
|Sensitivity||< 1 pg/mL|
|Predicted Homology Based on Immunogen Sequence||No detectable cross-reactivity with any other cytokine.|
|Datasheets/Manuals||Click here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.|
|Target Reference||1. Brenner, D. A.; O'Hara, M.; Angel, P.; Chojkier, M.; Karin, M. Prolonged activation of JUN and collagenase genes by
tumour necrosis factor-alpha. Nature 337: 661-663, 1989.
2. Nedwin, G. E.; Naylor, S. L.; Sakaguchi, A. Y.; Smith, D.; Jarrett-Nedwin, J.; Pennica, D.; Goeddel, D. V.; Gray, P. W.
Human lymphotoxin and tumor necrosis factor genes: structure, homology and chromosomal localization. Nucleic
Acids Res. 13: 6361-6373, 1985.
3. Broudy, V. C.; Kaushansky, K.; Segal, G. M.; Harlan, J. M.; Adamson, J. W. Tumor necrosis factor type alpha
stimulates human endothelial cells to produce granulocyte/macrophage colony-stimulating factor. Proc. Nat. Acad.
Sci. 83: 7467-7471, 1986.
4. Wang, A. M.; Creasey, A. A.; Ladner, M. B.; Lin, L. S.; Strickler, J.; Van Arsdell, J. N.; Yamamoto, R.; Mark, D. F.
Molecular cloning of the complementary DNA for human tumor necrosis factor. Science 228: 149-154, 1985.
|Specificity||There is no detectable cross-reactivity with other relevant proteins.|
|Official Gene Full Name||tumor necrosis factor|
|Alias Symbols||Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNFSF2|
|NCBI Gene Id||21926|
|Protein Name||Tumor necrosis factor|
|Description of Target||Tumor necrosis factor-alpha (TNF-alpha, or TNF) is secreted by macrophages in response to inflammation, infection and cancer.1 Human Tumor Necrosis Factor (TNF) and Lymphotoxin (TNF-beta) are cytotoxic proteins which have similar biological activities and share 30% amino acid homology.2 TNF-alpha is produced by monocytes, which can stimulate endothelial cells to produce the multilineage growth factor granulocyte-macrophage colony-stimulating factor and extend the role of this immunoregulatory protein to the regulation of hematopoiesis in vitro.3 TNF is a soluble protein that causes damage to tumor cells but has no effect on normal cells. Human TNF has been purified to apparent homogeneity as a 17.3-kilodalton protein from HL-60 leukemia cells and has showed cytotoxic and cytostatic activities against various human tumor cell lines. The human TNF cDNA is 1585 base pairs in length and encodes a protein of 233 amino acids. The mature protein begins at residue 77, leaving a long leader sequence of 76 amino acids.4 TNF-alpha has been mapped to human chromosome 6.2|
|Protein Accession #||NP_001265530.1|
|Nucleotide Accession #||NM_001278601.1|
|Tissue Tool||Find tissues and cell lines supported by DNA array analysis to express TNF.|
|RNA Seq||Find tissues and cell lines supported by RNA-seq analysis to express TNF.|
- Reconstitution & Storage Instructions
- Western Blotting/Immunoblotting (WB/IB) Protocol
- Immunohistochemistry (IHC) Protocol
- Immunocytochemistry (ICC) Protocol
- Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol
- Blocking Peptide Competition Protocol (BPCP)
- Immunoprecipitation (IP) Protocol
- Antibody Array (AA) Protocol
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