Mouse Autophagy Related Protein 7 (ATG7) ELISA Kit
Principle of the Assay
The microtiter plate provided in this kit has been pre-coated with an antibody specific to ATG7. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to ATG7. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain ATG7, 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 ATG7 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
For Use with serum, plasma, and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Target Information
E1-like activating enzyme involved in the 2 ubiquitin-like systems required for cytoplasm to vacuole transport (Cvt) and autophagy. Activates ATG12 for its conjugation with ATG5 as well as the ATG8 family proteins for their conjugation with phosphatidylethanolamine. Both systems are needed for the ATG8 association to Cvt vesicles and autophagosomes membranes. Facilitates LC3-I lipidation with phosphatidylethanolamine to form LC3-II which is found on autophagosomal membranes (By similarity). Required for autophagic death induced by caspase-8 inhibition. Required for mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Modulates p53/TP53 activity to regulate cell cycle and survival during metabolic stress. Also plays a key role in the maintenance of axonal homeostasis, the prevention of axonal degeneration, the maintenance of hematopoietic stem cells, the formation of Paneth cell granules, as well as in adipose differentiation. Plays a role in regulating the liver clock and glucose metabolism by mediating the autophagic degradation of CRY1 (clock repressor) in a time-dependent manner (PubMed:29937374).
GENE ID | 74244 |
SWISS PROT | Q9D906 |
SYNONYMS |
APG7L; GSA7; Ubiquitin-like modifier-activating enzyme ATG7; ATG12-activating enzyme E1 ATG7; Ubiquitin-activating enzyme E1-like protein |
Materials Supplied
Kit Components | 96 Wells Quantity/Size |
---|---|
Pre-coated, ready-to-use 96-well strip plate | 1 plate |
Plate sealer for 96 wells | 2 |
Standard |
2 tubes |
Diluent buffer | 1 bottle |
Detection Reagent A | 1 bottle |
Detection Reagent B | 1 bottle |
TMB Substrate | 1 tube |
Stop Solution | 1 tube |
Wash Buffer (30 ℅ concentrate) | 1 tube |
Product data sheet | 1 copy |
Storage
Storage | The TMB Substrate, Wash Buffer (30X concentrate), and the Stop Solution should be stored at 4°C upon receipt, while the other items should be stored at -20°C. |
Performance Characteristics
REPEATABILITY |
Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level ATG7 were tested 20 times on one plate, respectively. |
SENSITIVITY | The minimum detectable dose was 0.053ng/mL. |
ASSAY RANGE | 0.156-10ng/mL |
SPECIFICITY | This assay has high sensitivity and excellent specificity for the detection of ATG7. No significant cross-reactivity or interference between ATG7 and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between ATG7 and all analogs, therefore, cross reactivity may still exist. |