KEY FEATURES
| Full Name | Mammalian Target of Rapamycin |
|---|---|
| Synonym | FRAP; FRAP1; FRAP2; MTOR; RAFT1; RAPT1; FK506 Binding Protein 12 Rapamycin Associated Protein; Mammalian Target Of Rapamycin; FKBP12-Rapamycin Complex-Associated Protein 1; Rapamycin and FKBP12 target |
| Assay Type | Sandwich |
| Reactivity | Human |
| Range | 0.31-20ng/mL |
| Sensitivity | 0.12ng/mL |
| Sample Type | Serum,plasma and other biological fluids |
| Sample Volume | 100μL |
| Detection Wavelength | OD450 |
| Transportation Temperature | 2-8°C |
| Specificity | The kit detected Human mTOR in the samples and no significant cross-species reactions were found |
| Microplate | 96-wells plate breakable into 12 x 8 wells strip |
TEST PRINCIPLE
| This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human mTOR. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human mTOR and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human mTOR, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of 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 Human mTOR. You can calculate the concentration of Human mTOR in the samples by comparing the OD of the samples to the standard curve. |
ELISA KIT COMPONENTS
Upon receipt, unpack promptly and store as recommended in the instructions.
| Components | Specifications | Storage and Notes |
|---|---|---|
| Micro Plate | 96T: 8 wells×12 strips 48T: 8 wells×6 strips | Unopened: -20°C, 12 months Unused: Put it back in the aluminum foil bag and seal it, store it at -20°C. |
| Reference Standard | 96T: 2 vials 48T: 1 vial | Unopened: -20°C, 12 months Please use freshly dissolved standards for each experiment. Discard any unused standards after dissolution. |
| Biotinylated Detection Ab Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C, 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| HRP Conjugate Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C(Protect from light), 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| Biotinylated Detection Ab Diluent | 14mL×1 | 2-8℃, 12 months |
| HRP Conjugate Diluent | 14mL×1 | 2-8℃, 12 months |
| Reference Standard & Sample Diluent | 20mL×1 | 2-8℃, 12 months |
| Washing Buffer Concentrate (25×) | 30mL×1 | 2-8℃, 12 months |
| Substrate Reagent(TMB) | 10mL×1 | 2-8°C(Protect from light),12 months |
| Stop Solution | 7mL×1 | 2-8°C/Room temperature |
ASSAY PROCEDURES
TYPICAL DATA
Human mTOR ELISA Standard Curve
Typical data are for reference only and curves should be replotted for each experiment. The Logistics function is recommended for fitting.
PRECISION
Intra-Assay Precision (Precision within an assay): Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays): Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 0.94 | 3.13 | 9.54 | 0.91 | 3.22 | 10.35 |
| Standard deviation | 0.05 | 0.17 | 0.51 | 0.05 | 0.17 | 0.35 |
| CV(%) | 5.84 | 5.32 | 5.33 | 5.36 | 5.17 | 3.39 |
RECOVERY
The recovery of Human mTOR spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum(n=8) | 86-100 | 91 |
| EDTA plasma (n=8) | 90-106 | 97 |
| Cell culture media (n=8) | 96-108 | 102 |
LINEARITY
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Human mTOR in various matrices were diluted with the Reference Standard & Sample Diluent to produce samples with values within the dynamic range of the assay.
Pathways associated with ELISA Kit
| Adaptive Immune System Pathway | Adaptive Immune System Pathway |
| CD28 Co-stimulation Pathway | CD28 Co-stimulation Pathway |
| CD28 Dependent PI3K/Akt Signaling Pathway | CD28 Dependent PI3K/Akt Signaling Pathway |
| Constitutive PI3K/AKT Signaling In Cancer Pathway | Constitutive PI3K/AKT Signaling In Cancer Pathway |
| Costimulation By The CD28 Family Pathway | Costimulation By The CD28 Family Pathway |
| DAP12 Interactions Pathway | DAP12 Interactions Pathway |
| DAP12 Signaling Pathway | DAP12 Signaling Pathway |
| Disease Pathway | Disease Pathway |
| Downstream Signaling Events Of B Cell Receptor (BCR) Pathway | Downstream Signaling Events Of B Cell Receptor (BCR) Pathway |
| Downstream Signal Transduction Pathway | Downstream Signal Transduction Pathway |
Diseases associated with PIA ELISA Kit
| Neoplasms | publications with PIA and Neoplasms |
| Insulin Resistance | publications with PIA and Insulin Resistance |
| Breast Neoplasms | publications with PIA and Breast Neoplasms |
| Neoplasms, Experimental | publications with PIA and Neoplasms, Experimental |
| Kidney Diseases | publications with PIA and Kidney Diseases |
| Neoplasm Invasiveness | publications with PIA and Neoplasm Invasiveness |
| Hypertrophy | publications with PIA and Hypertrophy |
| Adenocarcinoma | publications with PIA and Adenocarcinoma |
| Necrosis | publications with PIA and Necrosis |
| Inflammation | publications with PIA and Inflammation |
Organs/Tissues associated with PIA ELISA Kit
| Kidney | publications with PIA and Kidney |
| Brain | publications with PIA and Brain |
| Bone | publications with PIA and Bone |
| Adipose Tissue | publications with PIA and Adipose Tissue |
| Bone Marrow | publications with PIA and Bone Marrow |
| Skin | publications with PIA and Skin |
| Heart | publications with PIA and Heart |
| Lung | publications with PIA and Lung |
| Lymph Node | publications with PIA and Lymph Node |
| Pancreas | publications with PIA and Pancreas |
