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Cyclic Nucleotide (cAMP and cGMP) Assays and Capture ELISA for Quantitative Analysis of Plasmodium falciparum Blood-stage Egress.
Upon rupture of Plasmodium falciparum (P. falciparum) schizonts in vitro (an occasion often known as egress), merozoites are launched into the tradition medium. The merozoites invade recent purple blood cells, a course of that entails shedding of a microneme protein known as apical membrane antigen-1 (AMA1) from the merozoite floor.
This shedding, which takes place even within the absence of invasion, is due to this fact a surrogate marker for the diploma of egress going down in a tradition, and could be measured utilizing a selected seize ELISA to quantify AMA1 ranges in tradition supernatants (Collins et al., 2013).
The assay makes use of a monoclonal antibody particular for AMA1 (known as 4G2dc1) (Kocken et al., 1998; Collins et al., 2009) to seize and immobilize the protein from tradition supernatants, then makes use of a selected rabbit polyclonal antiserum to detect the immobilized antigen.
A phosphatase-conjugated goat anti-rabbit antibody is lastly used to quantify the binding of the second antibody. Egress is completely dependent upon the exercise of a parasite cGMP-dependent protein kinase, PKG, and so is influenced by ranges of intracellular cGMP (Collins et al., 2013). That is regulated by the interaction between guanylate cyclases and phosphodiesterases. The latter enzymes may additionally degrade cAMP, so it might even be informative to measure intracellular cAMP ranges.
An enzyme-linked immunosorbent assay (ELISA) for guanosine 3′,5′-cyclic monophosphate (cGMP) in human plasma and urine utilizing monoclonal antibody.
An ELISA for cGMP in human plasma and urine utilizing a monoclonal antibody is described. The monoclonal antibody was raised in opposition to succinyl cGMP conjugated to human serum albumin. The conjugate was adsorbed to the ELISA plate, giving an immobilized antigen strategy which simplifies subsequent assay procedures. As little as 1.56 fmol/nicely of each plasma and urinary cGMP is measurable.
Recoveries of added cGMP in plasma and urine had been from 97% to 105%. Intra-assay coefficients of variation had been lower than 7.0% for plasma and seven.1% for urine samples. Inter-assay coefficients of variation for plasma and urine samples had been lower than 9.9% and 9.5%, respectively. The values obtained by ELISA correlated nicely with these by radioimmunoassay (RIA) (plasma: r = 0.96, n = 50; urine: r = 0.98, n = 60).
The impact of salmon calcitonin in opposition to glutamate-induced cytotoxicity within the C6 cell line and the roles the inflammatory and nitric oxide pathways play’
Current proof has proven that salmon calcitonin (sCT) has constructive results on the nervous system. Nevertheless, its impact and mechanisms on glutamate-induced cytotoxicity are nonetheless unclear. The present experiment was designed to look at the impact of sCT on glutamate-induced cytotoxicity in C6 cells, involving the inflammatory and nitric oxide stress pathways. The examine used the C6 glioma cell line.
4 cell teams had been ready to judge the impact of sCT on glutamate-induced cytotoxicity. The management group was with none remedy. Cells within the glutamate group had been handled with 10 mM glutamate for 24 h. Cells within the sCT group had been handled with numerous concentrations (3, 6, 12, 25, and 50 µg/mL) of sCT for 24 h. Cells within the sCT + glutamate group had been pre-treated with numerous concentrations of sCT for 1 h after which uncovered to glutamate for 24 h. The cell viability was evaluated with an XTT assay.
Nuclear issue kappa b (NF-kB), tumor necrosis issue alpha (TNF-α), interleukin-6 (IL-6), neuronal nitric oxide synthase (nNOS), nitric oxide (NO), cyclic guanosine monophosphate (cGMP), caspase-3, and caspase-9 ranges within the cells had been measured by ELISA kits. Apoptosis was detected by movement cytometry technique. sCT in any respect concentrations considerably improved the cell viability in C6 cells after glutamate-induced cytotoxicity (p < 0.001).
Furthermore, sCT considerably decreased the degrees of NF-kB (p < 0.001), TNF-α, and IL-6 ranges (p < 0.001). sCT additionally decreased nNOS, NO, and cGMP ranges (P < 0.001). Moreover, it decreased the apoptosis charge and elevated the live-cell charge within the movement cytometry (P < 0.001).
In conclusion, sCT has protecting results on glutamate-induced cytotoxicity in C6 glial cells by inhibiting inflammatory and nitric oxide pathways. sCT could possibly be a helpful supportive agent for individuals with neurodegenerative signs.
Correlation evaluation between C natriuretic peptide and being pregnant final result
C-type natriuretic peptide (CNP) performs a key function in feminine copy and is said to oocyte high quality. This examine analyzed the connection of CNP with being pregnant final result to offer a brand new indicator of being pregnant final result. Follicular fluids had been collected from 158 sufferers present process the IVF/ICSI process on the Middle for Reproductive Medication, Tongji Medical Faculty, Huazhong College of Science and Expertise.
CNP and cGMP ranges in human follicular fluids had been examined by ELISA. Then, the distribution patterns of CNP and NPR-B from GV oocytes to blastocysts in mice had been examined by confocal microcopy. Lastly, CNP was added to the fertility or embryo improvement medium to watch fertility charge and the event of the embryo. CNP ranges in follicular fluids from nonpregnant ladies had been considerably larger than these in follicular fluids from pregnant ladies.
A powerful constructive correlation between CNP and cGMP concentrations in human follicular fluids was noticed. Each CNP and NPR-B had been expressed within the plasma of cells at totally different levels from GV to blastocyst. CNP may improve the 2-cell charge of embryos and the speed of blastocysts when added to both fertility tradition medium or embryo tradition medium.
In a phrase, CNP in human follicular fluid may predict the being pregnant final result of IVF sufferers, and the mechanism of CNP in follicular fluid is said to the standard of oocyte or embryo competence and will promote the event of embryos.
Silencing Matrix Metalloproteinase 9 Exerts a Protecting Impact on Astrocytes After Oxygen-Glucose Deprivation and Is Correlated With Suppression of aquaporin-4
Cerebral ischemia, adopted by mind edema, could be life-threatening. It has been broadly reported that matrix metalloproteinase-9 (MMP-9) and aquaporin-4 (AQP4) have distinguished roles within the improvement of mind edema. Nevertheless, the precise mechanisms by which MMP-9 and AQP4 affect mind edema are usually not absolutely understood.
On this examine, astrocytes had been subjected to oxygen-glucose deprivation (OGD) /reperfusion (OGD/R) harm, an in vitro mannequin of Ischemia/reperfusion (I/R). Cell viability was evaluated via the measurement of LDH launch. The expression of MMP-9 and AQP4 additionally had been measured by qPCR and western blot. Subsequently, we knocked out the MMP-9 gene utilizing MMP-9 siRNA. AQP4 and its gene expression, and the LDH launch charge had been measured utilizing ELISA, Western blotting, and RT-PCR.
We additionally assessed cAMP-dependent protein kinase (PKA), cGMP-dependent protein kinase (PKG), protein kinase C (PKC), and Ca2+/calmodulin-dependent protein kinase II (CaMK II) in MMP-9 knockout astrocytes. All measurements had been carried out with or with out an OGD/R problem. OGD/reperfusion enhanced LDH launch ranges, and in addition elevated MMP-9 and AQP4 expression in astrocytes.
Monkey cGMP ELISA Kit |
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| EMKG0010 | Abclonal | 96Tests | EUR 625.2 |
Rabbit cGMP ELISA Kit |
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| ERTG0010 | Abclonal | 96Tests | EUR 625.2 |
Canine cGMP ELISA Kit |
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| ECG0010 | Abclonal | 96Tests | EUR 625.2 |
Bovine cGMP ELISA Kit |
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| EBG0010 | Abclonal | 96Tests | EUR 625.2 |
Porcine cGMP ELISA Kit |
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| EPG0010 | Abclonal | 96Tests | EUR 625.2 |
Chicken cGMP ELISA Kit |
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| ECKG0010 | Abclonal | 96Tests | EUR 625.2 |
Anserini cGMP ELISA Kit |
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| EAG0010 | Abclonal | 96Tests | EUR 625.2 |
cGMP ELISA Kit (OKEH02546) |
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| OKEH02546 | Aviva Systems Biology | 96 Wells | EUR 872.4 |
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Description: Description of target: Cyclic guanosine monophosphate (cGMP) is a cyclic nucleotide derived from guanosine triphosphate (GTP). cGMP acts as a second messenger much like cyclic AMP. Its most likely mechanism of action is activation of intracellular protein kinases in response to the binding of membrane-impermeable peptide hormones to the external cell surface. Guanylate cyclase (GC) catalyzes cGMP synthesis. This enzyme converts GTP to cGMP. In turn, peptide hormones such as the atrial natriuretic factor activate membrane-bound GC, while soluble GC is typically activated by nitric oxide to stimulate cGMP synthesis.;Species reactivity: All;Application: ;Assay info: Assay Methodology: Quantitative Competitive ELISA;Sensitivity: 5.8 pg/mL |
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cGMP ELISA Kit (Colorimetric) |
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| STA-505 | Cell Biolabs | 96 assays | EUR 706.8 |
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Description: Cyclic GMP (cGMP) is an important regulatory molecule in the GPCR signaling cascade. Our cGMP ELISA Kits provide a highly sensitive method to measure low levels of cyclic GMP in a variety of sample types. |
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cGMP ELISA Kit (Colorimetric) |
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| STA-505-5 | Cell Biolabs | 5 x 96 assays | EUR 2558.4 |
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Description: Cyclic GMP (cGMP) is an important regulatory molecule in the GPCR signaling cascade. Our cGMP ELISA Kits provide a highly sensitive method to measure low levels of cyclic GMP in a variety of sample types. |
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cGMP ELISA Kit (Chemiluminescent) |
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| STA-506 | Cell Biolabs | 96 assays | EUR 706.8 |
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Description: Cyclic GMP (cGMP) is an important regulatory molecule in the GPCR signaling cascade. Our cGMP ELISA Kits provide a highly sensitive method to measure low levels of cyclic GMP in a variety of sample types. |
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cGMP ELISA Kit (Chemiluminescent) |
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| STA-506-5 | Cell Biolabs | 5 x 96 assays | EUR 2558.4 |
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Description: Cyclic GMP (cGMP) is an important regulatory molecule in the GPCR signaling cascade. Our cGMP ELISA Kits provide a highly sensitive method to measure low levels of cyclic GMP in a variety of sample types. |
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Guinea Pig cGMP ELISA Kit |
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| EGG0010 | Abclonal | 96Tests | EUR 625.2 |
OKEH02546-96W - cGMP ELISA Kit |
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| OKEH02546-96W | Aviva Systems Biology | 96Wells | EUR 600 |
OKAU00058-1PLATE - cGMP ELISA Kit |
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| OKAU00058-1PLATE | Aviva Systems Biology | 1plate | EUR 400 |
OKAU00058-5PLATE - cGMP ELISA Kit |
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| OKAU00058-5PLATE | Aviva Systems Biology | 5plate | EUR 1469 |
OKAU00018-1PLATE - cGMP ELISA Kit |
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| OKAU00018-1PLATE | Aviva Systems Biology | 1plate | EUR 450 |
OKAU00018-5PLATE - cGMP ELISA Kit |
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| OKAU00018-5PLATE | Aviva Systems Biology | 5plate | EUR 1829 |
cGMP ELISA Kit| Rat Cyclic GMP ELISA Kit |
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| EF017622 | Lifescience Market | 96 Tests | EUR 826.8 |
cGMP(Cyclic GMP)ELISA Kit |
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| MBS2516175-10x96Tests | MyBiosource | 10x96Tests | EUR 3265 |
cGMP(Cyclic GMP)ELISA Kit |
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| MBS2516175-24Tests | MyBiosource | 24Tests | EUR 225 |
cGMP(Cyclic GMP)ELISA Kit |
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| MBS2516175-48Test | MyBiosource | 48Test | EUR 350 |
cGMP(Cyclic GMP)ELISA Kit |
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| MBS2516175-5x96Test | MyBiosource | 5x96Test | EUR 1655 |
cGMP(Cyclic GMP)ELISA Kit |
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| MBS2516175-96Tests | MyBiosource | 96Tests | EUR 395 |
cGMP(Cyclic GMP) ELISA Kit |
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| YPJ1147-48wellsplate | UpingBio | 48 wells plate | EUR 180 |
cGMP(Cyclic GMP) ELISA Kit |
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| YPJ1147-96wellsplate | UpingBio | 96 wells plate | EUR 220 |
Cyclic Guanosine Monophosphate (cGMP) ELISA Kit |
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| abx156669-100g | Abbexa | 100 µg | EUR 3325 |
Cyclic Guanosine Monophosphate (cGMP) ELISA Kit |
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| 20-abx156669 | Abbexa |
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Cyclic Guanosine Monophosphate (cGMP) ELISA Kit |
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| abx156669-200g | Abbexa | 200 µg | EUR 6300 |
Cyclic Guanosine Monophosphate (cGMP) ELISA Kit |
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| abx156669-50g | Abbexa | 50 µg | EUR 700 |
Cyclic Guanosine Monophosphate (cGMP) ELISA Kit |
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| abx576586-96tests | Abbexa | 96 tests | EUR 801.6 |
Cyclic Guanosine Monophosphate (cGMP) ELISA Kit |
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| DLR-cGMP-Ge-48T | DL Develop | 48T | EUR 681.6 |
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Description: A competitive inhibition quantitative ELISA assay kit for detection of Cyclic Guanosine Monophosphate (cGMP) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. |
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Silencing the MMP-9 gene decreased LDH launch ranges, and in addition was related to decreased AQP4 expression. The expression of PKC, however not PKA, PKG, or CaMK II, was decreased. This examine revealed that OGD/reperfusion may trigger cell harm in vitro. MMP-9 silencing protected astrocytes from hypoxic insult, and the protecting impact could also be enhanced by the downregulation of AQP4 expression. In conclusion, downregulating MMP-9 expression could also be helpful for the prevention and remedy of mind ischemia.
