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Figure 2: Assay Overview
Figure 4. cAMP Standard Curve. The provided cAMP standard containing 5 mM cAMP (5,000 pmol/µL) was used in this experiment. Seven serial dilutions wer...
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cAMP HTS Immunoassay Kit, 384 well (Chemiluminescent)
Description:
cAMP HTS Immunoassay Kit, 384 well (Chemiluminescent)
Trade Name:
Upstate (Millipore)
Background Information:
Cyclic AMP (cAMP, adenosine 3´, 5´-cyclic monophosphate) is a nucleotide which acts as a key second messenger in multiple signal transduction pathways [1]. It is synthesized from ATP by the action of adenylate cyclase, and is inactivated by hydrolysis to 5´-AMP by the actions of phosphodiesterases [2]. All receptors that act via cAMP are coupled to a stimulatory G protein, which activates adenylate cyclase upon ligand binding [3]. Many different drugs, neurotransmitters and hormones exert their cellular effects by modulating adenylate cyclase activity and thus raising or lowering intracellular cAMP concentrations [3,4].
cAMP regulates many cellular functions, such as metabolism, cell growth and differentiation, gene transcription, ion transport and ion channel function [3,4]. These cAMP effects, mediated primarily by cAMP-dependent protein kinase (PKA), result in cAMP being responsible for the regulation of many physiological processes, including cardiovascular, endocrine, neuronal, glandular, kidney, and immune functions, as well as general metabolism [5-11]. Consequently, agents which increase or decrease intracellular cAMP levels are of major interest in drug discovery [6,12].
Millipore's cAMP HTS immunoassay kit, 384 well is a competitive immunoassay for quantitative chemiluminescence-based detection of cAMP in cell lysates and supernatants, which has been optimized for use in high throughput screening applications. The assay is reproducible and robust, with a Z’ value of 0.7 and a S/B ratio of 92.4. Results can be obtained in as little as 1.5 hrs. Sensitivity of the assay is 1.07 fmol/well, and the assay has a large dynamic range, detecting cAMP concentrations from 0.0003 pmol/well to 3,000 pmol/well. The polyclonal antibody used is highly specific for cAMP and shows minimal cross-reactivity with other nucleotides. The kit provides two ready to use 384 well pre-coated assay microplates and all reagents required to perform the assay.
cAMP regulates many cellular functions, such as metabolism, cell growth and differentiation, gene transcription, ion transport and ion channel function [3,4]. These cAMP effects, mediated primarily by cAMP-dependent protein kinase (PKA), result in cAMP being responsible for the regulation of many physiological processes, including cardiovascular, endocrine, neuronal, glandular, kidney, and immune functions, as well as general metabolism [5-11]. Consequently, agents which increase or decrease intracellular cAMP levels are of major interest in drug discovery [6,12].
Millipore's cAMP HTS immunoassay kit, 384 well is a competitive immunoassay for quantitative chemiluminescence-based detection of cAMP in cell lysates and supernatants, which has been optimized for use in high throughput screening applications. The assay is reproducible and robust, with a Z’ value of 0.7 and a S/B ratio of 92.4. Results can be obtained in as little as 1.5 hrs. Sensitivity of the assay is 1.07 fmol/well, and the assay has a large dynamic range, detecting cAMP concentrations from 0.0003 pmol/well to 3,000 pmol/well. The polyclonal antibody used is highly specific for cAMP and shows minimal cross-reactivity with other nucleotides. The kit provides two ready to use 384 well pre-coated assay microplates and all reagents required to perform the assay.
Key Applications:
ELISA
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Quality Assurance:
Routinely evaluated by ELISA.
Kit or Assay Type:
Detection Kits
Components:
- 1. White 384-Well anti-Rabbit Coated Plate: (Part No. CS200427) Two 384-well immuno-plates pre-coated with anti-Rabbit polyclonal antibody, sealed in a foil pouch.
- 2. Rabbit anti-cAMP Antibody: (Part No. CS200428) One 20 μL vial.
- 3. cAMP Standard: (Part No. 2004073) One 2 mL bottle.
- 4. cAMP Alkaline Phosphatase Conjugated Tracer: (Part No. 2004072) One 20 μL vial.
- 5. 2x Assay Diluent: (Part No. CS200419) One 125 mL bottle.
- 6. 5x Wash Buffer: (Part No. CS200445) One 125 mL bottle.
- 7. Lysis Buffer: (Part No. CS200416) One 100 mL bottle.
- 8. 6x Alkaline Phosphatase Substrate: (Part No. CS200430) One 8 mL bottle.
- 9. Plate Sealers: (Part No. 2007518) 6 each.
Detection Methods:
Chemiluminescent
Packaging:
Kit capacity: 768 wells
Alternate Names:
cyclic AMP ELISA
Materials Required but Not Delivered:
1. Pipettors & tips capable of accurately measuring 10-1000 μL
2. Plate shaker (optional)
3. Automated Liquid Handler (optional)
4. Plate Washer (optional)
5. Luminometer capable of reading 384-well microplates.
6. Test tubes for standard and sample dilutions
7. Mechanical vortex
2. Plate shaker (optional)
3. Automated Liquid Handler (optional)
4. Plate Washer (optional)
5. Luminometer capable of reading 384-well microplates.
6. Test tubes for standard and sample dilutions
7. Mechanical vortex
Analytes Available:
cAMP