SynonymsDisintegrin and metalloproteinase domain-containing protein 17, ADAM 17, TNF-alpha convertase, TNF-alpha-converting enzyme, CD156b, Adam17, Tace
ReactivityMouse
SKU:
MOEB1148
UPC:
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9Z0F8
ELISA Type:
Sandwich
Synonyms:
Disintegrin and metalloproteinase domain-containing protein 17, ADAM 17, TNF-alpha convertase, TNF-alpha-converting enzyme, CD156b, Adam17, Tace
Mouse Disintegrin and metalloproteinase domain-containing protein 17 (Adam17) ELISA Kit
The Mouse ADAM17 ELISA Kit is specifically designed for the precise measurement of disintegrin and metalloproteinase domain-containing protein 17 (ADAM17) levels in mouse serum, plasma, and cell culture supernatants. This kit provides exceptional sensitivity and accuracy, ensuring dependable and consistent results, making it an invaluable tool for a variety of research purposes.ADAM17 is a critical enzyme that plays a pivotal role in various cellular processes, including cell signaling, inflammation, and immune response regulation.
Dysregulation of ADAM17 has been implicated in numerous diseases, such as cancer, inflammatory disorders, and cardiovascular conditions, underscoring its significance as a biomarker for studying these diseases and exploring potential therapeutic interventions.Overall, the Mouse ADAM17 ELISA Kit offers researchers a reliable and efficient method for quantifying ADAM17 levels in mouse samples, facilitating further investigations into the role of this enzyme in health and disease.
Product Name:
Mouse Disintegrin and metalloproteinase domain-containing protein 17 (Adam17) ELISA Kit
SKU:
MOEB1148
Size:
96T
Target:
Mouse Disintegrin and metalloproteinase domain-containing protein 17 (Adam17)
Synonyms:
TNF-alpha convertase, TNF-alpha-converting enzyme, CD156b, ADAM 17, Tace
Assay Type:
Sandwich
Detection Method:
ELISA
Reactivity:
Mouse
Detection Range:
0.078-5ng/mL
Sensitivity:
0.043ng/mL
Intra CV:
Provided with the Kit
Inter CV:
Provided with the Kit
Linearity:
Provided with the Kit
Recovery:
Provided with the Kit
Function:
Cleaves the membrane-bound precursor of TNF-alpha to its mature soluble form. Responsible for the proteolytical release of soluble JAM3 from endothelial cells surface. Plays a role in the proteolytic processing of ACE2 (By similarity). 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, growth hormone receptor, MUC1 and the amyloid precursor protein. Acts as an activator of Notch pathway by mediating cleavage of Notch, generating the membrane-associated intermediate fragment called notch extracellular truncation (NEXT).
Uniprot:
Q9Z0F8
Sample Type:
Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:
Natural and recombinant mouse Disintegrin and metalloproteinase domain-containing protein 17
Sub Unit:
Interacts with MAD2L1, MAPK14 and MUC1.
Research Area:
Neurosciences
Subcellular Location:
Isoform Short Secreted
Storage:
Please see kit components below for exact storage details
Note:
For research use only
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.Protein type: Motility/polarity/chemotaxis; Protease; Membrane protein, integral; EC 3.4.24.86Cellular Component: cell surface; focal adhesion; membrane; integral to plasma membrane; cytoplasm; apical plasma membrane; extracellular region; plasma membrane; integral to membrane; intercellular junction; actin cytoskeleton; lipid raftMolecular Function: integrin binding; peptidase activity; protein binding; interleukin-6 receptor binding; zinc ion binding; metallopeptidase activity; hydrolase activity; metalloendopeptidase activity; metal ion binding; Notch binding; SH3 domain binding; PDZ domain bindingBiological Process: cell motility involved in cell locomotion; T cell differentiation in the thymus; response to lipopolysaccharide; proteolysis; G1/S-specific positive regulation of cyclin-dependent protein kinase activity; positive regulation of epidermal growth factor receptor activity; positive regulation of cell proliferation; negative regulation of interleukin-8 production; germinal center formation; cell adhesion; epidermal growth factor receptor signaling pathway; response to drug; spleen development; Notch signaling pathway; response to high density lipoprotein stimulus; membrane protein ectodomain proteolysis; positive regulation of transforming growth factor beta receptor signaling pathway; Notch receptor processing; positive regulation of cell motility; positive regulation of cell growth; positive regulation of chemokine production; regulation of mast cell apoptosis; PMA-inducible membrane protein ectodomain proteolysis; B cell differentiation; response to hypoxia; cell adhesion mediated by integrin; positive regulation of protein amino acid phosphorylation; negative regulation of transforming growth factor beta receptor signaling pathway; negative regulation of apoptosis; positive regulation of cell migration
disintegrin and metalloproteinase domain-containing protein 17 isoform 2
NCBI Synonym Full Names:
a disintegrin and metallopeptidase domain 17
NCBI Official Symbol:
Adam17
NCBI Official Synonym Symbols:
Tace; CD156b
NCBI Protein Information:
disintegrin and metalloproteinase domain-containing protein 17; TNF-alpha convertase; TNF-alpha converting enzyme; a disintegrin and metalloproteinase domain 17
UniProt Protein Name:
Disintegrin and metalloproteinase domain-containing protein 17
Disintegrin and metalloproteinase domain-containing protein
UniProt Gene Name:
Adam17
UniProt Entry Name:
ADA17_MOUSE
Component
Quantity (96 Assays)
Storage
ELISA Microplate (Dismountable)
8×12 strips
-20°C
Lyophilized Standard
2
-20°C
Sample Diluent
20ml
-20°C
Assay Diluent A
10mL
-20°C
Assay Diluent B
10mL
-20°C
Detection Reagent A
120µL
-20°C
Detection Reagent B
120µL
-20°C
Wash Buffer
30mL
4°C
Substrate
10mL
4°C
Stop Solution
10mL
4°C
Plate Sealer
5
-
Other materials and equipment required:
Microplate reader with 450 nm wavelength filter
Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
Incubator
Deionized or distilled water
Absorbent paper
Buffer resevoir
*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.
Step
1.
Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2.
Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3.
Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4.
Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5.
Repeat the wash process for five times as conducted in step 3.
6.
Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7.
Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8.
Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9.
After experiment, store all reagents according to the specified storage temperature respectively until their expiry.
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
Sample Type
Protocol
Serum
If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma
Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid
Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant
Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates
Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates
The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates
Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk
Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.
