DAB Substrate Solution for HRP

Abstract Rooted cuttings of Cotoneaster dammeri Schneid ‘Skogholm’ and seedlings of Rudbeckia fulgida Ait. ‘Goldsturm’ were potted into 3.8 liter (4 qt) containers in a pine bark:sand (8:1 by vol) substrate incorporated with 3.5 g (0.12 oz) N per container provided by one of the following five controlled-release fertilizers (CRFs): Meister 21N–3.5P–11.1K (21–7–14), Osmocote 24N–2.0P–5.6K (24–4–7), Scotts 23N–2.0P–6.4K (23–4–8), Sustane 5N–0.9P–3.3K (5–2–4) or Woodace 21N–3.0P–9.5K (21–6–12). Two hundred ml (0.3 in), 400 ml (0.6 in), 800 ml (1.1 in) or 1200 ml (1.7 in) of water was applied once daily (single) or in two equal applications with a 2 hr interval between applications (cyclic). Substrate solutions were collected from containers of cotoneaster 15, 32, 45, 60, 74, 90, 105, and 119 days after initiation (DAI). Irrigation efficiency [(water applied − water leached) ÷ water applied] was determined on the same days. Cyclic application improved irrigation efficiency at 800 ml (1.1 in) and 1200 ml (1.7 in) ≈ 27% compared to a single application. Irrigation efficiencies averaged over the season were 95%, 84%, 62%, and 48% for cotoneaster and 100%, 90%, 72%, and 51% for rudbeckia at 200 ml (0.3 in), 400 ml (0.6 in), 800 ml (1.1 in) and 1200 ml (1.7 in), respectively. NH4-N and NO3-N and PO4-P concentrations in substrate solution decreased with increasing irrigation volume regardless of CRF. Substrate NH4-N concentration decreased throughout the season with most CRFs below 5 mg/liter by 90 DAI. CRFs mainly affected substrate NH4-N and NO3-N concentrations when irrigated with 200 ml (0.3 in) or 400 ml (0.6 in). Substrate NH4-N, NO3-N, and PO4-P solution concentrations were similar for all CRFs at irrigation volume of 1200 ml (1.7 in). Osmocote, Scotts, and Woodace maintained relatively constant substrate solution levels of PO4-P through 60 DAI. By 90 DAI, substrate PO4-P levels were similar regardless of irrigation volume or CRF. Substrate PO4-P concentrations were never in the recommended range of 5 to 10 mg/liter when irrigated with 800 ml (1.1 in) or 1200 ml (1.7 in) regardless of CRF. Solution pH remained in the recommended range of 5.0 to 6.0 for all irrigation volumes and CRFs throughout the entire study with the exception of Sustane.

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Enterohemorrhagic Escherichia coli Colony Check Assay for the Identification of Serogroups O26, O45, O103, O111, O121, O145, and O157 Colonies Isolated on Plating Media

Journal of Food Protection ◽

10.4315/0362-028x.jfp-13-555 ◽

2014 ◽

Vol 77 (7) ◽

pp. 1212-1218 ◽

Cited By ~ 1

Author(s):

BURTON BLAIS ◽

MYLÈNE DESCHÊNES ◽

GEORGE HUSZCZYNSKI ◽

MARTINE GAUTHIER

Keyword(s):

Escherichia Coli ◽

Shiga Toxin ◽

High Throughput Screening ◽

Enterohemorrhagic Escherichia Coli ◽

Test Results ◽

Bacterial Strains ◽

Enrichment Broth ◽

Substrate Solution ◽

E Coli ◽

Assay Performance

A simple immunoenzymatic enterohemorrhagic Escherichia coli (EHEC) colony check (ECC) assay was developed for the presumptive identification of priority EHEC colonies isolated on plating media from enrichment broth cultures of foods. With this approach, lipopolysaccharide extracted from a colony is spotted on the grid of a polymyxin-coated polyester cloth strip, and bound E. coli serogroup O26, O45, O103, O111, O121, O145, and O157 antigens are subsequently detected by sequential reactions with a pool of commercially available peroxidase-conjugated goat antibodies and tetramethylbenzidine substrate solution. Each strip can accommodate up to 15 colonies, and test results are available within 30 min. Assay performance was verified using colonies from a total of 73 target EHEC isolates covering the range of designated priority serogroups (all of which were reactive), 41 nontarget E. coli isolates including several nontarget Shiga toxin–producing E. coli serogroups (all unreactive), and 33 non–E. coli strains (all unreactive except two bacterial strains possessing O-antigenic structures in common with those of the priority EHEC). The ECC assay was reactive with target colonies grown on several types of selective and nonselective plating media designed for their cultivation. These results support the use of the ECC assay for high-throughput screening of colonies isolated on plating media for detecting priority EHEC strains in foods.

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Electrical conductivity and pH of the substrate solution in gerbera cultivars under fertigation

Horticultura Brasileira ◽

10.1590/s0102-05362013000300003 ◽

2013 ◽

Vol 31 (3) ◽

pp. 356-360 ◽

Cited By ~ 7

Author(s):

Fernanda Ludwig ◽

Dirceu M Fernandes ◽

Poliana RD Mota ◽

Roberto L Villas Bôas

Keyword(s):

Electrical Conductivity ◽

Experimental Design ◽

Nutrient Uptake ◽

Plant Nutrition ◽

Substrate Solution ◽

Plant Toxicity ◽

Paulo State ◽

Ph Values ◽

Nutritive Solution ◽

Golden Yellow

The quality and the profitability on floriculture are intimately linked to the adequate plant nutrition. In the present research we aimed to evaluate the electrical conductivity (EC) and pH of the substrate solution on four different gerbera cultivars subjected to fertigation, with two nutritive solutions. The experiment was carried out in a greenhouse, from May to July 2006, on the Universidade Estadual Paulista, Botucatu, São Paulo state, Brazil. The experiment was carried out under an experimental design of random blocks, in 4×2 factorial arrangement, with four Gerbera cultivars (Cherry, Golden Yellow, Salmon Rose and Orange) and two nutritive solution concentrations: 0.92 and 1.76 dS m-1 EC) during the vegetative stage, and 1.07 and 2.04 dS m-1 during the reproductive stage (S1 and S2, respectively). The nutrients were applied through fertigation, manually performed every day. The EC and pH values of the substrate solution were evaluated weekly, using the 'pourthru' method. Orange and Cherry cultivars had, respectively, the highest and the lowest electrical conductivity of the substrate solution, and Cherry was the most efficient on the nutrient uptake. The solution S2 showed a trend to accumulate salts in the substrate, but without visual symptoms of plant toxicity, leading to the lowest pH values. The 'pourthru' method was efficient when compared to the 1:2 method and can be adopted for substrate solution analysis in gerbera culture.

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Standardization of substrate and buffer concentrations for in-vivo nitrate reductase activity in Adenanthera microsperma leaves

Indian Journal of Forestry ◽

10.54207/bsmps1000-2015-5dbcb9 ◽

2015 ◽

Vol 38 (4) ◽

pp. 309-311

Author(s):

Priyanshu Sharma ◽

S.P. Chaukiyal ◽

Meenu Sengar

Keyword(s):

Nitrate Reductase ◽

Phosphate Buffer ◽

Nitrate Reductase Activity ◽

Reductase Activity ◽

Buffer Solution ◽

Substrate Solution ◽

Substrate Concentrations

The combination of different substrate concentrations (0.05M, 0.10M, 0.15M, 0.20M and 0.25M, KNO3) with different pH of phosphate buffer (0.10 M and 0.20 M, KH2PO4 of the pH 7.0, 7.5, 7.6, 7.7, and 7.8) solutions were tried for in-vivo nitrate reductase activity of Adenanthera microsperma leaves. Maximum nitrate reductase activity was observed in the combination of buffer solution (0.20M KH2PO4) having pH 7.7 and substrate solution 0.20 M concentration.

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An accelerated enzyme immunoassay for human choriogonadotropin in urine, involving reflow of specimen through capillary tubes.

Clinical Chemistry ◽

10.1093/clinchem/33.4.498 ◽

1987 ◽

Vol 33 (4) ◽

pp. 498-501 ◽

Cited By ~ 2

Author(s):

H M Chandler ◽

S A Fuller ◽

C H Nakagawa ◽

P A Nagainis ◽

J G Hurrell

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Enzyme Immunoassay ◽

Test Specimen ◽

Capillary Tube ◽

Test Procedure ◽

Beta Subunit ◽

Sensitive Assay ◽

Substrate Solution ◽

Human Choriogonadotropin

Abstract A very rapid and sensitive assay for human choriogonadotropin (hCG) has been developed involving two beta-subunit-specific monoclonal antibodies. In the assay the test specimen is passed backward and forward (reflow) through a monoclonal-antibody-coated capillary tube for 1 min, then incubated for 1 min with a second monoclonal antibody conjugated to urease (EC 3.5.1.5). After addition of a urease substrate solution, 10 int. units of hCG per liter can be detected visually within 5 min, which compares very favorably with other currently available hCG assay procedures. Advantages of the reflow/capillary tube assay system and optimization of the test procedure are discussed.

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d-Luciferin Substrate Solution (7.5 mg/mL)

Cold Spring Harbor Protocols ◽

10.1101/pdb.rec079673 ◽

2015 ◽

Vol 2015 (1) ◽

pp. pdb.rec079673

Keyword(s):

Substrate Solution

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Detection of thymic surface antigens and radioactive labeling of mouse lymphoid cells.

Journal of Histochemistry & Cytochemistry ◽

10.1177/25.11.334963 ◽

1977 ◽

Vol 25 (11) ◽

pp. 1269-1274 ◽

Cited By ~ 1

Author(s):

D Allison ◽

K Bose ◽

J L Hill

Keyword(s):

Specific Surface ◽

Simultaneous Detection ◽

Surface Antigens ◽

Lymphoid Cells ◽

Rabbit Serum ◽

Target Cells ◽

Tissue Specific ◽

Substrate Solution ◽

Antigen Assay

A method for the permanent and simultaneous detection of tissue-specific surface antigens and internal radioactive labeling of mouse lymphoid cells is described. Target cells were first reacted with a mouse-derived "antithymocyte serum", incubated with peroxidase-conjugated rabbit serum against mouse immunoglobulins and placed in a substrate solution that leads to staining of the antigen-positive cells. Radioactive labeling was demonstrated by autoradiography performed after the antigen assay. More than 90% of antigen-positive thymocytes could be specifically stained in the assay without staining of similarly treated antigen-negative cells. Autoradiographic grains could be detected over both antigen-positive and antigen-negative cells.

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Fluorometric Method for Measuring Serum Lipase Activity

Clinical Chemistry ◽

10.1093/clinchem/21.12.1788 ◽

1975 ◽

Vol 21 (12) ◽

pp. 1788-1790 ◽

Cited By ~ 3

Author(s):

Bernd Rietz ◽

George G Guilbault

Keyword(s):

Enzyme Activity ◽

Olive Oil ◽

Lipase Activity ◽

Enzymatic Reaction ◽

Fluorometric Method ◽

Reaction Volume ◽

Serum Lipase ◽

Fluorescent Indicator ◽

Substrate Solution ◽

Emulsified Olive Oil

Abstract We describe a method for determining lipase (triacylglycerol acyl-hydrolase, EC 3.1.1.3) activity in serum. Emulsified olive oil in tris(hydroxymethyl)aminomethane buffer is used as substrate. The course of the enzymatic reaction is followed by measuring the decrease in fluorescence with time of a fluorescent indicator, 4-methylumbelliferone, added to the substrate, caused by the change in the pH of the substrate solution resulting from the reaction. All measurements are performed at 37 °C. The reaction volume is about 2.3 ml, in a Pyrex cuvette. Change of fluorescence and of enzyme activity are linearly related in the range of 77.8 to 389.0 U/liter. An assay can be done in as little as 3 to 5 min, with excellent precision.

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