INSR ELISA Kit (Rat) (OKCD02586)

Catalog No: OKCD02586

Size:96WELLS

Rat INSR ELISA kit standard curve

Product Info

• Predicted Species Reactivity: Rat|Rattus norvegicus
• Application: Enzyme-linked Immunosorbent assay-Sandwich
• ELISA Kit Detection Method: Colorimetric
• ELISA Kit Duration: 3h
• ELISA Kit Linearity:

  Sample	1:2	1:4	1:8	1:16
  Serum (n=5)	78-94%	79-101%	83-102%	90-97%
  EDTA Plasma (n=5)	83-92%	98-105%	83-105%	81-101%
  Heparin Plasma (n=5)	88-95%	80-104%	87-95%	93-101%

• ELISA Kit Principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Insulin Receptor (INSR). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Insulin Receptor (INSR). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Insulin Receptor (INSR), 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 Insulin Receptor (INSR) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
• ELISA Kit Range: 0.78-50ng/mL
• ELISA Kit Recovery:

  Matrix	Recovery range (%)	Average(%)
  Serum (n=5)	81-97	92
  EDTA Plasma (n=5)	82-89	86
  Heparin Plasma(n=5)	80-101	80

• ELISA Kit Reproducibility: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Insulin Receptor (INSR) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Insulin Receptor (INSR) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
• ELISA Kit Component:
  • Anti-INSR Microplate: 96 Wells (12 x 8 Well strips)
  • INSR Lyophilized Standard: 2
  • 100X Biotinylated INSR Detector Antibody: 1 x 120 uL
  • 100X Avidin-HRP Conjugate: 1 x 120 uL
  • Standard Diluent: 1 x 20 mL
  • Detector Antibody Diluent: 1 x 12 mL
  • Conjugate Diluent: 1 x 12 mL
  • 30X Wash Buffer: 1 x 20 mL
  • Stop Solution: 1 x 6 mL
  • TMB Substrate: 1 x 9 mL
• Reconstitution and Storage: 2°C to 8°C|-20°C
• Sample Type: Serum, plasma, tissue homogenates and other biological fluids
• Sensitivity: 0.34 ng/mL
• Specificity: This assay has high sensitivity and excellent specificity for detection of Insulin Receptor (ISR). No significant cross-reactivity or interference between Insulin Receptor (ISR) and analogues was observed.
• Assay Info: Assay Methodology: Quantitative Sandwich ELISA

Target Info

• Gene Symbol: INSR
• Alias Symbols: Insulin receptor, IR
• Protein Name: Insulin receptor
• Description of Target: Receptor tyrosine kinase which mediates the pleiotropic actions of insulin. Binding of insulin leads to phosphorylation of several intracellular substrates, including, insulin receptor substrates (IRS1, 2, 3, 4), SHC, GAB1, CBL and other signaling intermediates. Each of these phosphorylated proteins serve as docking proteins for other signaling proteins that contain Src-homology-2 domains (SH2 domain) that specifically recognize different phosphotyrosine residues, including the p85 regulatory subunit of PI3K and SHP2. Phosphorylation of IRSs proteins lead to the activation of two main signaling pathways: the PI3K-AKT/PKB pathway, which is responsible for most of the metabolic actions of insulin, and the Ras-MAPK pathway, which regulates expression of some genes and cooperates with the PI3K pathway to control cell growth and differentiation. Binding of the SH2 domains of PI3K to phosphotyrosines on IRS1 leads to the activation of PI3K and the generation of phosphatidylinositol-(3, 4, 5)-triphosphate (PIP3), a lipid second messenger, which activates several PIP3-dependent serine/threonine kinases, such as PDPK1 and subsequently AKT/PKB. The net effect of this pathway is to produce a translocation of the glucose transporter SLC2A4/GLUT4 from cytoplasmic vesicles to the cell membrane to facilitate glucose transport. Moreover, upon insulin stimulation, activated AKT/PKB is responsible for: anti-apoptotic effect of insulin by inducing phosphorylation of BAD; regulates the expression of gluconeogenic and lipogenic enzymes by controlling the activity of the winged helix or forkhead (FOX) class of transcription factors. Another pathway regulated by PI3K-AKT/PKB activation is mTORC1 signaling pathway which regulates cell growth and metabolism and integrates signals from insulin. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 thereby activating mTORC1 pathway. The Ras/RAF/MAP2K/MAPK pathway is mainly involved in mediating cell growth, survival and cellular differentiation of insulin. Phosphorylated IRS1 recruits GRB2/SOS complex, which triggers the activation of the Ras/RAF/MAP2K/MAPK pathway. In addition to binding insulin, the insulin receptor can bind insulin-like growth factors (IGFI and IGFII). When present in a hybrid receptor with IGF1R, binds IGF1 (By similarity).
• Uniprot ID: P15127
• Protein Accession #: NP_058767.2
• Protein Size (# AA): 1383