CREB Regulated Transcription Coactivator 1 (CRTC1) BioAssay(TM) ELISA Kit (Mouse)

Catalog Number: USB-589166
Article Name: CREB Regulated Transcription Coactivator 1 (CRTC1) BioAssay(TM) ELISA Kit (Mouse)
Biozol Catalog Number: USB-589166
Supplier Catalog Number: 589166
Alternative Catalog Number: USB-589166-96
Manufacturer: US Biological
Category: Kits/Assays
The CREB Regulated Transcription Coactivator 1 (CRTC1) BioAssay(TM) ELISA Kit utilizes the sandwich enzyme immunoassay technique for the in vitro quantitative measurement of mouse CRTC1 in tissue homogenates, cell lysates, cell culture supernates and other biological fluids. Detection Range: 0.312-20ng/ml Sensitivity: 0.129ng/ml Specificity: This assay has high sensitivity and excellent specificity for detection of CREB Regulated Transcription Coactivator 1 (CRTC1). No significant cross-reactivity or interference between CREB Regulated Transcription Coactivator 1 (CRTC1) and analogues was observed. Precision: Intra-Assay %CV: <10% Inter-Assay %CV: <12% Test Principle: The microplate provided in this kit has been pre-coated with an antibody specific to CRTC1. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to CRTC1. 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 CRTC1, 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 CRTC1 in the samples is then determined by comparing the O.D. of the samples to the standard curve. Kit Components: Pre-coated, ready to use 96-well strip plate Standard, 2x vials Detection Reagent A, 1x120ul Detection Reagent B, 1x120ul TMB Substrate, 1x9ml Wash Buffer (30X): 1x20ml Standard Diluent, 1x20ml Assay Diluent A, 1x12ml Assay Diluent B, 1x12ml Stop Solution, 1x6ml Storage and Stability: Store Microtiter Strips, Standard, Detection Reagents and Positive Control at -20C. After reconstitution of Positive Control, store at 4C and use within 5 days. Store all the other components at 4C. Unused kit is stable for 6 months after receipt. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap. Assay Procedure Summary: 1. Prepare all reagents, samples and standards. 2. Add 100ul standard or sample to each well. Incubate 1 hour at 37C. 3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37C. 6. Aspirate and wash 5 times. 7. Add 90ul Substrate Solution. Incubate 10-20 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately.