All tissue culture reagents were from Invitrogen (Carlsbad, CA, USA), unless otherwise specified. Fetal bovine serum (FBS) was from Gemini Bio-products (Woodland, CA, USA). Aβ25–35, Aβ35–25, Aβ1–42, and Aβ42–1 peptides, as well as nitro-L-arginine methyl ester (L-NAME), were from Sigma-Aldrich (St. Louis, MO, USA). Monoclonal and polyclonal anti-eNOS and anti-HSP90 antibodies were from BD-Transduction Laboratories (San Diego, CA, USA). The polyclonal antibody for phospho-eNOS (Ser 1179) was from Invitrogen (Grand Island, NY, USA). Polyclonal and monoclonal antibodies for anti-Akt and phospho-Akt (Ser473) were purchased from Cell Signaling (Danvers, MA, USA). Protein A/G agarose beads were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The ECL chemiluminescence detection assay as well as the peroxidase-conjugated anti-mouse/anti-rabbit IgG were from Amersham Biosciences (Piscataway, NJ, USA). The cGMP enzyme-immunoassay kit was from Cayman Chemical Co. (Ann Arbor, MI, USA). The blots were reprobed after stripping using a stripping solution from Pierce (Rockford, IL, USA).
Cgmp enzyme immunoassay kit
Manufactured by Cayman Chemical
Sourced in United States
The CGMP enzyme immunoassay kit is a laboratory tool designed for the quantitative measurement of cyclic guanosine monophosphate (cGMP) levels in various sample types. The kit utilizes a competitive enzyme immunoassay principle to determine the concentration of cGMP in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Fura 2 am 2 acetoxymethyl, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488 conjugated fibrinogen, by Thermo Fisher Scientific (2 mentions)
Thromboxane b2 assay kit, by Cayman Chemical (2 mentions)
Aβ35 25, by Merck Group (1 mentions)
Nitro l arginine methyl ester l name, by Merck Group (1 mentions)
Aβ25 35, by Merck Group (1 mentions)
Aβ1 42, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Gemini Bio (1 mentions)
Protein a g agarose bead, by Santa Cruz Biotechnology (1 mentions)
Anti phospho pi3k, by Cell Signaling Technology (1 mentions)
Anti phospho p38 mapk, by Cell Signaling Technology (1 mentions)
Atp assay kit, by Cayman Chemical (1 mentions)
Anti phospho cpla2, by Cell Signaling Technology (1 mentions)
Anti phospho akt, by Cell Signaling Technology (1 mentions)
Mouse monoclonal to cd62p p selectin antibody, by BioLegend (1 mentions)
Antiphospho vasp, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Thrombin, by Chrono-Log (1 mentions)
U46619, by Chrono-Log (1 mentions)
10 protocols using cgmp enzyme immunoassay kit
1
Investigating Amyloid-Beta Pathways
2
Renal Perfusion Dynamics and cGMP Production
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Perfusate flow from kidneys isolated as described above was calculated by collection and gravimetric determination of the venous effluent. After establishing a constant perfusion pressure (100 mmHg), perfusate flow rates stabilised within 12–15 min. Stock solutions of the indicated drugs were added to the perfusate. For evaluation, the amount of venous effluent was normalised to the perfusate flow under control conditions or after application of 30 μM ODQ as indicated in the respective graphs to exclude differences in baseline perfusion between the preparations. For determination of renal cGMP production, venous effluent was collected over a period of 1 min during four intervals along the study and its cGMP concentration was determined after acetylation of samples using a cGMP enzyme immunoassay Kit (Cat# 581021, Cayman Chemical, Hamburg, Germany). The cGMP secretion was calculated by multiplying cGMP concentration and perfusate flow.
3
Ovarian cGMP Determination Protocol
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Ovarian tissues were solubilized in 200 µl of 5% trichloroacetic acid (TCA). After removing TCA, the samples were used for cGMP assay by a cGMP enzyme immunoassay kit obtained from Cayman Chemicals (Ann Arbor, MI, USA).
4
Mulberroside C Signaling Pathway Analysis
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Mulberroside C (Figure 1 ) was provided by ChemFaces (Wuhan, China). Mouse monoclonal to CD62P (P-selectin) antibody was purchased from Biolegend (San Diego, CA, USA). Fura 2-AM (2-acetoxymethyl) and Alexa Fluor 488-conjugated fibrinogen were obtained from Invitrogen (Eugene, OR, USA). The BCA protein assay kit was obtained from Pierce Biotechnology (Rockford, IL, USA). The serotonin ELISA kit was provided by Labor Diagnostika Nord GmbH and Co. (Nordhorn, Germany). Collagen, U46619, and thrombin were purchased from Chrono-Log Co. (Havertown, PA, USA). The thromboxane B2 assay kit, cAMP, cGMP enzyme immunoassay kit, and ATP assay kit were obtained from Cayman Chemical (Ann Arbor, MI, USA). Cell signaling (Beverly, MA, USA) supplied antiphospho-p38MAPK, antiphospho-ERK (1/2), antiphospho-VASP (Ser157), antiphospho-VASP (Ser239), antiphospho-cPLA2 (Ser505), antiphospho-PI3K (Tyr458), antiphospho-Akt (Ser473), antiphospho-inositol-3-phosphate receptor type I (Ser1756), antiphospho-PLCγ2 (Tyr759), anti-β-actin, and antirabbit secondary antibodies.
5
Quantifying cGMP and ET-1 in cell pellets
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Cell pellets (n=6) were acidified with 0.6% trifluoroacetic acid to give a 10% w/v homogenate, which was centrifuged at 2,000 × g for 15 min at 4°C. The supernatant was washed and dried under a stream of nitrogen at 60°C, and the dried extract was dissolved in a 20 µL assay buffer prior to analysis. cGMP level was determined using a cGMP enzyme immunoassay kit (Cayman Chemical, Ann Arbor, MI, USA), and the secreted ET-1 was measured using an enzyme immunoassay kit (Biomedica Group, Wien, Austria).
6
Cyclic GMP Measurement in Cardiomyocytes
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Isolated ventricular cardiomyocytes (cultured for 48 h to ensure similar conditions as cells used for FRET imaging) and HEK293 cells were stimulated in the presence and absence of 50 nM SNAP, 100 µM SNAP and/or 300 nM CNP (as indicated) for 10 min. Cyclic GMP levels were measured using a cGMP enzyme immunoassay kit (Cayman Chemical Company, Ann Arbor, MI, USA) as previously described36 (link).
7
Cyclic GMP Quantification in COCs
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COCs from eCG-primed mice were collected after different drug treatments for 2 h. 100 COC samples per group were solubilized in 200 µL HCl (0.1 M) on ice for 10 min, and then were thawed and centrifuged at 12,000 × g for 5 min. The supernatant was collected to a tube and dried in an oven at 60 °C. The levels of cGMP were determined using a cGMP enzyme immunoassay kit obtained from Cayman Chemicals (Ann Arbor, MI, USA).
8
Platelet Activation and Signaling Assays
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Derrone was purchased from ChemFaces (Wuhan, China). Collagen was purchased from Chrono-Log Co. (Havertown, PA, USA). Fura 2-AM (2-acetoxymethyl) and Alexa Fluor 488-conjugated fibrinogen were obtained from Invitrogen (Eugene, OR, USA). Serotonin ELISA kit was purchased from Labor Diagnostika Nord GmbH and Co. (Nordhorn, Germany). Bicinchoninic acid protein assay kit was purchased from Pierce Biotechnology (IL, USA). Cayman chemical (Ann Arbor, MI, USA) offered thromboxane B2 assay kit, cAMP, cGMP enzyme immunoassay kit, and U46619. Cell Signaling (Beverly, MA, USA) supplied all antibodies. The adhesion kit (fibronectin coated) was obtained from Cell Biolabs (San Diego, CA, USA).
9
Quantifying cGMP in HT1080 Cells
10
cGMP Measurement in Capacitated Sperm
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The capacitated spermatozoa of 100 μL (1 × 107 cells/mL) were incubated without or with 1 nM NPPC for 20 min, and then collected and solubilized in 100 μL of 0.1 M HCl on ice for 15 min. The samples were thawed and centrifuged for 5 min at 12,000× g, and the supernatant was collected and dried in an oven at 60 °C for cGMP assay. The levels of cGMP were determined by a cGMP-enzyme immunoassay kit (Cayman Chemical, Ann Arbor, MI, USA) according the manufacturer’s instruction.
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