aCella™ - cGMP
Cyclic GMP Chemiluminescent Immunoassay Kit
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| Key Benefits: |
- Ultra-sensitive - Measures as little as < 1 fmol cGMP per sample.
- Flexible - One kit for Extra- and Intra-Cellular cGMP quantification.
- Convenient - Stable, 4 C liquid reagents.
- Ready-to-Use - Complete kit with validated Cell Lysis Reagent Included.
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| Introduction to aCella - cGMP |
Cyclic GMP is a critical and multifunctional secondary messenger present at levels typically 10-100 fold lower than cAMP in most tissues. Intracellular cGMP is formed by the action of the enzyme guanylate cyclase on GTP and degraded through phosphodiesterase hydrolysis. Guanylate cyclases (GC) are either soluble or membrane bound. Soluble GCs are nitric oxide responsive, whereas membrane bound GCs respond to hormones such as acetylcholine, insulin and oxytocin. Other chemicals like serotonin and histamine also cause an increase in cGMP levels. Cycle GMP regulates cellular composition through cGMP-dependent kinase, cGMP-dependent ion channels or transporters and through its hydrolytic degradation by phosphodiestererase.
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| Assay Principle |
Cell Technology's aCella – cGMP Chemiluminescent Immunoassay kit measures cGMP present in lysed cells, EDTA, plasma, urine, saliva and culture media samples. The kit is unique in that all samples and standards are diluted into an acidic Sample Diluent, which contain special additives and stabilizers, for cGMP measurement. Acidified samples of cGMP are stable and endogenous phosphodiesterases are inactived in the Sample Diluent. After an overnight incubation at 4 C, the plate is washed and the chemiluminescent substrate is added. The substrate generates light that is detected in a multi-label microtiter plate reader capable of reading luminescence. |
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| References |
- Domek-Lopacinska, K. and Strosznajder, JB. “Cyclic GMP metabolism and its role in brain physiology “ (2005) J Physiol Pharmacol 56 Suppl 2, 15-34.
- Lucas, K.A. et al. “Guanylyl cyclases and signaling by cyclic GMP “ (2000) Pharmacol Rev 52: 375-414.
- Ashman, DF, et al., “Isolation of adenosine 3’, 5’-monophosphate and guanosine 3’, 5’-monophosphate from rat urine.“ (1963), Biochem Biophys Res Comm, 11: 330-4.
- Potter LR, Abbey-Hosch S, and Dickey DM. “Natriuretic peptides, their receptors, and cyclic guanosine monophosphate-dependent signaling functions“. (2006) Endocr Rev 27: 47-72.
- Waldman, SA and Murad, F., “Cyclic GMP synthesis and function “ (1987) Pharmacol Revs, 39: 163-197.
- Tremblay J, Gerzer R, and Hamet P., “Cyclic GMP in cell function”. (1988) , Adv. 2nd Messanger & Phosphoprotein Res., 22: 319-383.
- Matsumoto T, Kobayashi T, and Kamata K “Phosphodiesterases in the vascular system.” (2003) J Smooth Muscle Res 39: 67-86.
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| Kit contents: |
| 1. | Coated White 96 Well Plates (Part# 90001): White plastic microtiter plate coated with donkey anti-donkey anti-sheep IgG. Store at 2-8 C upon arrival. |
| 2. | Cyclic GMP Standard (Part# 90002): 1 vial (70 uL) cyclic GMP at 640 pmol/mL in a special stabilizing solution. Store at 2-8 C upon arrival. |
| 3. | Cyclic GMP Antibody (Part# 90003): 1 bottle (3 mL) mouse monoclonal antibody specific for cyclic GMP. Store at 2-8 C upon arrival. |
| 4. | Cyclic GMP Conjugate Concentrate (Part# 90004): 1 vial (150 uL) cyclic GMP-peroxidase conjugate concentrate in a special stabilizing solution. Store at 2-8 C upon arrival. |
| 5. | Conjugate Diluent (Part# 90005): 1 bottle (3 mL) Diluent containing special stabilizers and additives. Store at 2-8 C upon arrival. |
| 6. | Sample Diluent (Part# 90006): 1 bottle (28 mL) diluent containing special stabilizers and additives. Ready to use. Store at 2-8 C upon arrival. WARNING: CAUSTIC. |
| 7. | Plate Primer (Part# 90007): 1 bottle (5 mL) neutralizing solution containing special stabilizers and additives. Store at 2-8 C upon arrival. |
| 8. | Acetic Anhydride (Part# 90008): 1 bottle (2 mL). Store at 2-8 C upon arrival. WARNING: Corrosive Lachrymator |
| 9. | Triethylamine (Part# 90009): 1 bottle (4 mL). Store at 2-8 C upon arrival. WARNING: Corrosive Lachrymator |
| 10. | Wash Buffer Concentrate (Part # 90010): 1 bottle (30 mL) 20x concentrate that must be diluted with deionized or distilled water. Store at 2-8 C upon arrival. |
| 11. | Substrate Solution A (Part # 90011): 1 bottle (6 mL). Upon arrival store at 2-8 C. |
| 12. | Substrate Solution B (Part # 90012): 1 bottle (6 mL). Upon arrival store at 2-8 C. |
| 13. | Plate Sealer (Part # 90013) |
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