cGMP Parameter Assay Kit

R&D Systems | Catalog # KGE003

Item in the process of being discontinued
R&D Systems
Discontinued Product
KGE003 has been discontinued. View all cGMP products.

Key Product Details

Assay Length

3.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Cell Lysates (100 µL), EDTA Plasma (100 µL), Saliva (100 µL), Urine (100 µL)

Sensitivity

3.06 pmol/mL

Assay Range

2.1-500 pmol/mL (Cell Culture Supernates, Cell Lysates, EDTA Plasma, Saliva, Urine)
View all cGMP ELISA Kits »

Product Summary for cGMP Parameter Assay Kit

The Parameter cGMP Immunoassay is a 3.5 hour competitive enzyme immunoassay designed to measure cGMP in cell culture supernates, plasma, saliva, urine, and cell lysates.

Product Specifications

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

Biotin

Species

Multi-Species

Specificity

Measures cyclic GMP levels in various samples

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity not tested.

Interference

No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, EDTA Plasma, Saliva, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 85.2 184 265 80.7 188 273
Standard Deviation 6.9 12.5 14.0 8.0 16.5 22.4
CV% 8.1 6.8 5.3 9.9 8.8 8.2

Cell Lysates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 57.0 72.0 178 52.0 67.0 195
Standard Deviation 2.2 4.0 9.5 4.0 4.4 12.7
CV% 3.9 5.6 5.3 7.7 6.6 6.5

Recovery for cGMP Parameter Assay Kit

The recovery of cGMP spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 100 91-108
Cell Lysates (n=1) 92 80-98
EDTA Plasma (n=4) 91 74-105
Saliva (n=4) 102 80-123
Urine (n=4) 90 77-98

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of cGMP were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Multi-species cGMP ELISA Linearity

Scientific Data Images for cGMP Parameter Assay Kit

Multi-species cGMP ELISA Cell Lysate Standard Curve

Multi-species cGMP ELISA Cell Lysate Standard Curve

Multi-species cGMP ELISA Cell Culture Supernate/Plasma/Saliva/Urine Standard Curve

Multi-species cGMP ELISA Cell Culture Supernate/Plasma/Saliva/Urine Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: cGMP

Guanosine 3, 5-cyclic monophosphate (cGMP) is formed via the action of guanylate cyclase on GTP. cGMP is present at levels typically 10-100 fold lower than cAMP in most tissues. A variety of stimuli including acetylcholine, insulin, oxytocin, serotonin, and histamine will cause an increase in cGMP levels. Various stimulators of guanylate cyclase (e.g. vasodilators) as well as peptides that relax smooth muscle also increase cGMP concentrations.

Long Name

Cyclic GMP

Product Documents for cGMP Parameter Assay Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for cGMP Parameter Assay Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for cGMP Parameter Assay Kit

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Protocols

View specific protocols for cGMP Parameter Assay Kit (KGE003):

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 150 µL Diluent to NSB wells & 100 µL Diluent to B0 wells
  3.   Add 150 µL of the appropriate diluent to the non-specific binding (NSB) wells and 100 µL of the appropriate diluent to the zero standard (B0) wells.
    4. 100 µL Standard, Control, or Sample
  4.   Add 100 µL of Standard, Control, or sample to the remaining wells.
    5. 50 µL Conjugate
  5.   Add 50 µL of Conjugate to each well.
    6. 50 µL Primary Antibody Solution
  6.   Add 50 µL of Primary Antibody Solution to each well (except the NSB wells).
  7.   Cover with a plate sealer, and incubate at room temperature for 3 hours on a horizontal orbital microplate shaker.
  8.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    10. 50 µL Stop Solution
  10. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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