cGMP HTS Immunoassay Kit, 384 well (Chemiluminescent)

cGMP HTS Immunoassay Kit, 384 well (Chemiluminescent)

Upstate (Millipore)

768 assays

Cyclic GMP (cGMP, Guanosine 3', 5'-cyclic monophosphate) is a nucleotide which acts as a key second messenger in multiple signal transduction pathways, and is found in most tissues at levels 10 to 100 times lower than cAMP [1]. cGMP is synthesized from guanosine triphosphate (GTP) by the action of guanylate cyclase, degraded by cyclic nucleotide phosphodiesterases, which hydrolyze cGMP into 5'-GMP. Levels of cGMP are increased by the action of certain hormones, such as acetylcholine, insulin and oxytocin, as well as by chemicals such as serotonin and histamine. cGMP can also be elevated by agents which stimulate guanylate cyclase, such as nitroprusside, nitroglycerin, sodium nitrate and nitric oxide [2].

Downstream effects of cGMP are mediated primarily by cGMP-gated ion channels, cGMP-dependent protein kinases and cGMP-regulated phosphodiesterases [3]. cGMP has become an important target for drug discovery because it regulates many cellular functions, including ion channel conductance, glycogenolysis, and cellular apoptosis [4, 5]. Significantly, it also relaxes smooth muscle tissues. In blood vessels, relaxation of vascular smooth muscles leads to vasodilation and increased blood flow. This effect of cGMP has led to the notable clinical success of Sildenafil (Viagra), which enhances the vasodilatory effects of cGMP within the corpus cavernosum by inhibiting a phosphodiesterase, and is widely used as a treatment for erectile dysfunction [6].

Millipore’s cGMP HTS immunoassay kit, 384 well is a competitive immunoassay for chemiluminescence-based detection of cGMP 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 47.8. Results can be obtained in as little as 1.5 hrs. Sensitivity of the assay is 2.7 fmol/well, and the assay has a large dynamic range, detecting cGMP concentrations from 0.00003 pmol/well to 300 pmol/well. The polyclonal antibody used is highly specific for cGMP 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.

ELISA

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.

Routinely evaluated by ELISA.

• 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-cGMP polyclonal antibody: (Part No. CS200432) One 40 μL vial.
• 3. cGMP Standard: (Part No. 2004762) One 2 mL bottle.
• 4.Alkaline Phosphatase cGMP Tracer Conjugate: (Part No. 2004760) One 20 μL vial.
• 5.2x cGMP Assay Diluent: (Part No. CS200433) 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.

Upstate

• Chemiluminescence
• Chemiluminescence

Kit capacity: 768 wells

cGMP HTS Immunoassay Kit, 384 well (Chemiluminescent)

cyclic GMP ELISA

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

cGMP