Nitrate Reductase in the Citrus Plant: Properties, Assay Conditions and Distribution within the Plant

1967 ◽  
Vol 20 (2) ◽  
pp. 500-506 ◽  
Author(s):  
A. Bar-Akiva ◽  
J. Sagiv
Keyword(s):  
Nitrate Reductase ◽  
Citrus Plant ◽  
Assay Conditions ◽  
Plant Properties

Effects of Environment on Nitrate Reductase Assays in Field-grown Cotton Leaves

1978 ◽  
Vol 14 (4) ◽  
pp. 317-323 ◽  
Author(s):  
G. C. Bate ◽  
M. E. R. Meakin ◽  
D. M. Oosterhuis
Keyword(s):  
Nitrate Reductase ◽  
Incubation Period ◽  
Nitrate Reductase Activity ◽  
Reductase Activity ◽  
Soil Nitrate ◽  
Light Conditions ◽  
Optimum Conditions ◽  
Assay Conditions

SUMMARYOptimum conditions are described for the in vivo measurement of nitrate reductase activity in field-grown cotton leaves. The incubation period depended on light conditions prior to sampling. Addition of 1 mM glycolate had the effect of increasing apparent nitrate reductase activity on most days tested, and followed the general rhythmical pattern exhibited by the enzyme assayed without glycolate. While the in vivo assay appears to correlate well with soil nitrate levels, the choice of assay conditions does not appear to be absolutely critical.

scholarly journals Characterization of nitrate reductase activity (NR) in foliar and radicular tissues of Physalis angulata L.: diurnal variations and protocol optimization

2019 ◽  
pp. 1120-1125
Author(s):  
Tamara Torres Tanan ◽  
Marilza Neves do Nascimento ◽  
Alismário Leite da Silva ◽  
David Santana Guimarães ◽  
Romeu da Silva Leite ◽  
...  
Keyword(s):  
Nitrate Reductase ◽  
Reductase Activity ◽  
Nitrate Assimilation ◽  
Assay Method ◽  
Physalis Angulata ◽  
Nr Activity ◽  
Assay Conditions

Nitrate reductase (NR) is the first enzyme in the nitrogen assimilation pathway. The determination of its activity requires modifications for each plant species. The goal of this work was to evaluate the variation of NR activity throughout the day and the optimization of assay conditions in foliar and radicular tissues of Physalis angulata. The analysis was done in plants cultivated in a hydroponic system at two months of age. The NR activity was based on the in vivo assay method. Enzyme activity was observed on leaf and root, indicating two sites of nitrate assimilation with a higher activity in the daylight in leaf. The NR activity in leaf was increased after 4h of luminosity. In the root, we observed a high activity during most of the day, especially in periods of higher solar radiation and temperature. To obtain the highest activity of NR in both tissues we suggest 1% n-propanol, 50mM of KNO3- in pH=7 phosphate buffer for 75 min incubation in water bath.

Difference of [Ca2+]i Movements in Platelets Stimulated by Thrombin and TRAP: the Involvement of αIIbβ3-Mediated TXA2 Synthesis

1998 ◽  
Vol 79 (06) ◽  
pp. 1184-1190 ◽  
Author(s):  
Yoshiaki Tomiyama ◽  
Shigenori Honda ◽  
Kayoko Senzaki ◽  
Akito Tanaka ◽  
Mitsuru Okubo ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Fibrinogen Binding ◽  
Adp Release ◽  
The Difference ◽  
Txa2 Receptor Antagonist ◽  
High Level ◽  
Inhibition Studies ◽  
Peak Increase ◽  
Second Peak ◽  
Assay Conditions

SummaryThis study investigated the difference of [Ca2+]i movement in platelets in response to thrombin and TRAP. The involvement of αIIbβ3 in this signaling was also studied. Stimulation of platelets with thrombin at 0.03 U/ml caused platelet aggregation and a two-peak increase in [Ca2+]i. The second peak of [Ca2+]i, but not the first peak was abolished by the inhibition of platelet aggregation with αIIbβ3 antagonists or by scavenging endogenous ADP with apyrase. A cyclooxygenase inhibitor, aspirin, and a TXA2 receptor antagonist, BM13505, also abolished the second peak of [Ca2+]i but not the first peak, although these regents did not inhibit aggregation. Under the same assay conditions, measurement of TXB2 demonstrated that αIIbβ3 antagonists and aspirin almost completely inhibited the production of TXB2. In contrast to thrombin-stimulation, TRAP caused only a single peak of [Ca2+]i even in the presence of platelet aggregation, and a high level of [Ca2+]i increase was needed for the induction of platelet aggregation. The inhibition of aggregation with αIIbβ3 antagonists had no effect on [Ca2+]i change and TXB2 production induced by TRAP. Inhibition studies using anti-GPIb antibodies suggested that GPIb may be involved in the thrombin response, but not in the TRAP. Our findings suggest that low dose thrombin causes a different [Ca2+]i response and TXA2 producing signal from TRAP. Endogenous ADP release and fibrinogen binding to αIIbβ3 are responsible for the synthesis of TXA2 which results in the induction of the second peak of [Ca2+]i in low thrombin- but not TRAP-stimulated platelets.

Antibody Screening Results for Anti-Nucleocapsid Antibodies Towards the Development of a SARS-CoV-2 Nucleocapsid Protein Antigen Detecting Lateral Flow Assay

2021 ◽  
Author(s):  
David Cate ◽  
Helen Hsieh ◽  
Veronika Glukhova ◽  
Joshua D Bishop ◽  
H Gleda Hermansky ◽  
...  
Keyword(s):  
Nucleocapsid Protein ◽  
Point Of Care ◽  
Lateral Flow ◽  
Screening Methods ◽  
Antibody Screening ◽  
Liquid Handling ◽  
Large Numbers ◽  
Accurate Performance ◽  
Further Development ◽  
Assay Conditions

<p></p><p>The global COVID-19 pandemic has created an urgent demand for large numbers of inexpensive, accurate, rapid, point-of-care diagnostic tests. Analyte-based assays are suitably inexpensive and can be rapidly mass-produced, but for sufficiently accurate performance they require highly optimized antibodies and assay conditions. We used an automated liquid handling system, customized to handle arrays of lateral flow immunoassay (LFA) tests in a high-throughput screen, to identify anti-nucleocapsid antibodies that will perform optimally in an LFA. We tested 1021 anti-nucleocapsid antibody pairs as LFA capture and detection reagents with the goal of highlighting pairs that have the greatest affinity for unique epitopes of the nucleocapsid protein of SARS-CoV-2 within the LFA format. In contrast to traditional antibody screening methods (e.g., ELISA, bio-layer interferometry), the method described here integrates real-time reaction kinetics with transport in, and immobilization directly onto, nitrocellulose. We have identified several candidate antibody pairs that are suitable for further development of an LFA for SARS-CoV-2.</p><p></p>

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