Overview

  • Product name
    cGMP Assay Kit - Direct Immunoassay
  • Sample type
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay type
    Quantitative
  • Sensitivity
    > 0.008 µM
  • Range
    0.008 µM - 2 µM
  • Product overview

    Abcam's cGMP Assay Kit (ab65356) provides a direct competitive immunoassay for sensitive and quantitative determination of cGMP level in biological samples. The kit utilizes the recombinant Protein G coated plate to anchor cGMP polyclonal antibody. cGMP-HRP conjugates directly competes with cGMP from samples for binding to the cGMP specific antibody on the plate. After incubation and washing, the amount of cGMP-HRP bound to plate can easily be determined by reading OD450nm. The intensity of OD450nm is inversely proportional to the concentration of cGMP in samples. The kit provides a new acetylation procedure that improves detection signal significantly. The kit can detect 0.04-10 pmol/well (0.008-2 µM) cGMP samples.
    Visit our FAQs page for tips and troubleshooting.

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Storage buffer
    Preservative: None
    Constituents: BSA
  • Components Identifier 100 tests
    10X cGMP Assay Buffer WM 1 x 25ml
    Acetylating Reagent A Violet 1 x 750µl
    Acetylating Reagent B Black 1 x 1.5ml
    Anti-cGMP pAb/BSA Red 1 vial
    cGMP-HRP/BSA Green 1 vial
    HRP Developer Amber 1 x 10ml
    Neutralizing Buffer NM 1 x 7.5ml
    Protein G Coated Plate 1 pack
    Standard cGMP (10 nmol) Yellow 1 vial
  • Research areas
  • Relevance
    Cyclic guanosine monophosphate (cGMP) is a cyclic nucleotide derived from guanosine triphosphate (GTP). It serves as a second messenger in a manner similar to that observed with cAMP. Peptide hormones, such as the natriuretic factors, activate receptors that are associated with membrane-bound guanylate cyclase (GC). Receptor activation of GC leads to the conversion of GTP to cGMP. Nitric oxide (NO) also stimulates cGMP production by activating soluble GC, perhaps by binding to the heme moiety of the enzyme. Similar to cAMP, cGMP mediates most of its intracellular effects through the activation of specific cGMP dependent protein kinases (PKG).
  • Alternative names
    • Cyclic GMP
    • Cyclic guanosine monophosphate
    • Guanosine 3’ 5’ cyclic monophosphate

Images

  • O.D.'s of human urine measured at various dilutions (+/- SD)

  • Standard curve showing mean of duplicates (+/-SD) with background readings subtracted

Protocols

References

This product has been referenced in:
  • Suganya N  et al. Establishment of pancreatic microenvironment model of ER stress: Quercetin attenuates ß-cell apoptosis by invoking nitric oxide-cGMP signaling in endothelial cells. J Nutr Biochem 55:142-156 (2018). Read more (PubMed: 29455095) »
  • Sarkar O  et al. Nitric oxide attenuates overexpression of Gia proteins in vascular smooth muscle cells from SHR: Role of ROS and ROS-mediated signaling. PLoS One 12:e0179301 (2017). Rat . Read more (PubMed: 28692698) »
See all 5 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Question
Answer

You can use a speedvac to dry down samples.

Read More

Answer



After adding the acetylating reagent mix, incubate at RT for 10 mins, then put the tubes in a laminar airflow hood or simply air-dry. 5ul will dry-up fairly quickly. For low amounts of cGMP, add 50-100ul of the assay buffer to dried tubes and then proceed with quantification.

Read More

Answer

Thank you for contacting us.
On page 7 the section b of assay protocol states "for suspension cells collect by centrifugation. Add 1 ml of 0.1M HCl for every 35 cm2 of surface area (e.g., 10 cm plate at 70% confluency is ˜110 cm2, so use ˜3.1 ml). Incubate at room temperature for 20 minutes on ice."
This means you need to centrifuge out the media. The volume to HCL can be calculated as per the instructions given.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question
Answer

Thank you for contacting us.

The protcol booklet has been updated with missing information. Please check.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Answer

Thank you for your recent telephone enquiry.

I can confirm that the Bradford assay can be used for protein quantifation with this kit.

The BCA protein assay is not compatible with the acidic buffer in the samples you would have prepared with this kit.

I hope this information has been useful for you. Please do not hesitate to let me know if you have any other questions.

Read More

Answer

Yes, the supernatant from this extraction can be stored at -80°C in aliquots for later analysis.

Read More

Question
Answer

We don’t have a recommended protocol for collection of blood for this kit. I would just suggest storing the blood on ice before processing it for serum/plasma collection.


Hope this information has been helpful for you. Please let me know if you have any other questions.

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up