Rat TACE (TNF alpha Converting Enzyme) ELISA Kit (RTES00720)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- ADAM metallopeptidase domain 17,ADAM17,TACE,tumor necrosis factor-alpha-converting enzyme,ADAM18, CD156B, CSVP, NISBD,
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||0.31-20 ng/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Rat TACE in samples. No significant cross-reactivity or interference between Rat TACE and analogues was observed.|
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat TACE. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat TACE and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat TACE, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat TACE. The concentration of Rat TACE in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||TACE: a type I membrane protein with disintegrin and metalloprotease activity. An ubiquitously expressed ectoenzyme of peptidase family M12B. Must be membrane anchored to cleave the different substrates. The cytoplasmic domain is not required for the this activity. Possesses a narrow endopeptidase specificity. Cleaves Pro-Leu-Ala-Gln-Ala-|-Val-Arg-Ser-Ser-Ser in the membrane-bound, 26-kDa form of tumor necrosis factor alpha (TNF-alpha) to its mature soluble form. Responsible for the proteolytic release of several other cell-surface proteins, including p75 TNF-receptor, interleukin 1 receptor type II, p55 TNF-receptor, transforming growth factor-alpha, L-selectin, and the amyloid precursor protein. Also involved in the activation of Notch pathway. Binds 1 zinc ion per subunit. Two splice variant isoforms have been described.|
|UniProt Protein Details:|
Protein type:EC 3. 4. 24. 86; Membrane protein, integral; Motility/polarity/chemotaxis; Protease
Chromosomal Location of Human Ortholog: 6q16
Cellular Component: actin cytoskeleton; apical plasma membrane; cell surface; cytoplasm; cytosol; extracellular space; focal adhesion; integral component of membrane; integral component of plasma membrane; intercellular junction; membrane; membrane raft; plasma membrane
Molecular Function:integrin binding; interleukin-6 receptor binding; metal ion binding; metalloendopeptidase activity; metallopeptidase activity; Notch binding; PDZ domain binding; SH3 domain binding
Biological Process: B cell differentiation; cell adhesion; cell adhesion mediated by integrin; cell motility; defense response to Gram-positive bacterium; epidermal growth factor receptor signaling pathway; germinal center formation; integrin-mediated signaling pathway; membrane protein ectodomain proteolysis; negative regulation of apoptosis; negative regulation of neuron projection development; negative regulation of transforming growth factor beta receptor signaling pathway; Notch receptor processing; Notch signaling pathway; PMA-inducible membrane protein ectodomain proteolysis; positive regulation of cell growth; positive regulation of cell migration; positive regulation of cell proliferation; positive regulation of cellular component movement; positive regulation of chemokine production; positive regulation of cyclin-dependent protein serine/threonine kinase activity involved in G1/S transition of mitotic cell cycle; positive regulation of epidermal growth factor-activated receptor activity; positive regulation of protein phosphorylation; positive regulation of T cell chemotaxis; positive regulation of transforming growth factor beta receptor signaling pathway; production of molecular mediator of acute inflammatory response; proteolysis; regulation of axon regeneration; regulation of mast cell apoptotic process; regulation of neuron migration; response to drug; response to high density lipoprotein stimulus; response to hypoxia; response to lipopolysaccharide; spleen development; T cell differentiation in thymus; wound healing, spreading of epidermal cells
|NCBI Summary:||expression occurs in response to oxygen glucose deprivation, may play a role in inhibition of apoptosis via induction of TNF-alpha release [RGD, Feb 2006]|
|NCBI GenInfo Identifier:||9945330|
|NCBI Gene ID:||57027|
|NCBI Accession:||NP_064702. 1|
|UniProt Related Accession:||Q9Z1K9|
|Molecular Weight:||93,017 Da|
|NCBI Full Name:||disintegrin and metalloproteinase domain-containing protein 17|
|NCBI Synonym Full Names:||ADAM metallopeptidase domain 17|
|NCBI Official Symbol:||Adam17|
|NCBI Official Synonym Symbols:||TACE|
|NCBI Protein Information:||disintegrin and metalloproteinase domain-containing protein 17|
|UniProt Protein Name:||Disintegrin and metalloproteinase domain-containing protein 17|
|UniProt Synonym Protein Names:||TNF-alpha convertase; TNF-alpha-converting enzyme; CD_antigen: CD156b|
|Protein Family:||Disintegrin and metalloproteinase domain-containing protein|
|UniProt Gene Name:||Adam17|
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat TACE were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat TACE were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||7.00||4.26||4.84||4.95||4.30||5.42|
The recovery of Rat TACE spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||93-108||100|
|Cell culture media (n=5)||89-99||94|
Samples were spiked with high concentrations of Rat TACE and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro ELISA Plate(Dismountable)||8 wells ×12 strips||-20°C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100×)||1 vial, 120 µL|
|Concentrated HRP Conjugate (100×)||1 vial, 120 µL||-20°C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4°C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25×)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4°C(shading light)|
|Stop Solution||1 vial, 10 mL||4°C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.