Post-nuclear supernatants were diluted and incubated for 60 min with (-)-[3H]TZ659 in a total volume of 150 μl at 25 °C in 96-well polypropylene plates (Fisher Scientific, Pittsburgh, PA). Each well contained 20 μg protein while the concentration of the radioligand ranged from 0.15 nM to 20 nM. Reactions were terminated by the addition of 100 μl of the assay buffer at 4 °C, then samples were harvested and filtered rapidly using a 96-well glass fiber filtration plate (Millipore, Billerica, MA) presoaked with 100 μl assay buffer for 1 hour. Each filter was washed with 5 × 200 μl assay buffer then transferred to a scintillation vial with 2 ml of scintillation fluid and counted on a Wallac 1450 MicroBeta TriLux liquid scintillation counter (Perkin Elmer, Boston, MA). Nonspecific binding was determined from samples which contained 20 μM (-)-vesamicol hydrochloride. Counts were normalized to mg protein in the sample. The equilibrium dissociation constant (Kd) and maximum number of binding sites (Bmax) were determined by nonlinear regression analysis of onesite saturation binding model using GraphPad Prism 5.0 (GraphPad Software, Inc., San Diego, CA). Three independent experiments were performed; the results are reported as mean ± standard deviation.
96 well polypropylene plate
Manufactured by Thermo Fisher Scientific
Sourced in United States, Canada, Denmark
The 96-well polypropylene plates are a type of laboratory equipment used for various applications in scientific research and analysis. These plates feature a standard 96-well format and are made of polypropylene, a durable and commonly used material in the laboratory environment. The plates provide a convenient and organized platform for performing multiple experiments or analyses simultaneously.
Automatically generated - may contain errors
Lab products found in correlation
96 well glass fiber filtration plate, by Merck Group (2 mentions)
Prism 5, by GraphPad (2 mentions)
Wallac 1450 microbeta trilux, by PerkinElmer (1 mentions)
Bioplex 2200 system, by Bio-Rad (1 mentions)
Vidas benchtop immunoanalyzer, by bioMérieux (1 mentions)
Vidas hiv duo quick, by bioMérieux (1 mentions)
Elecsys anti sars cov 2 s rbd, by Roche (1 mentions)
Elecsys anti sars cov 2, by Roche (1 mentions)
Cobas e411 analyser, by Roche (1 mentions)
Specimen diluent, by Ortho Clinical Diagnostics (1 mentions)
Synergy h1, by Agilent Technologies (1 mentions)
Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Licor odyssey fc imaging system, by Thermo Fisher Scientific (1 mentions)
Bio dot dot blot apparatus, by Bio-Rad (1 mentions)
13 protocols using 96 well polypropylene plate
1
Radioligand Binding Assay Protocol
2
Biofilm Dispersal of S. Typhimurium
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Biofilms of S. Typhimurium ATCC14028 were set up as above using overnight cultures of the pathogen diluted 1:100 in the CFA medium in wells of 96-well polypropylene plates (Fisher, Waltham, MA, USA). Plates with bacteria were incubated for 24 hours at 37°C inside a Ziploc bag. Upon completion of the incubation, the medium with planktonic bacteria was removed and 10nmolL−1 of Molsidomine or MAHMA nonoate were added to the wells with biofilms. As controls, BPS alone was used. Plates were incubated at 22°C for 24 hours. Upon completion of the incubation, planktonic cells were removed, wells were washed twice with PBS and 200 μL of SaniDade 12.0 (BioSafe System, Hartford, CT, USA) diluted as per manufacturer’s recommendations and were loaded into the wells. The disinfectant was incubated for 10 minutes, after the incubation time, biofilm dispersal was measured by staining the remaining biofilms with 1% crystal violet in ethanol, as described previously (Merritt et al. [2005 (link)]; O’Toole and Kolter [1998 (link)]). 12 replicas for each experiment were done.
3
Radioligand Saturation Binding Assay
4
SARS-CoV-2 IgG Antibody DBS Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DBS specimens were tested using the BioPlex 2200 SARS-CoV-2 IgG assay (Bio-Rad Laboratories, Hercules, CA) at the NLHRS (Public Health Agency of Canada, Winnipeg, Canada) according to the manufacturer’s instructions coupled with BioPlex 2200 SARS-CoV-2 IgG Calibrators diluted in BioPlex 2200 Wash Buffer (Bio-Rad) using a 1:8 ratio. DBS specimens were punched (1 × 6 mm) using a semi-automated BSD600 Ascent puncher (BSD Robotics) into 400 μL 96-well polypropylene plates (ThermoFisher Scientific). DBS punches were eluted in 130 μL of DPBS containing 0.5% BSA and 0.05% Tween 20 overnight at 4 °C without agitation. Afterwards, plates were incubated at room temperature for 30 min with agitation (400 RPM) and 100 μL of DBS eluate was transferred into 2 mL microtubes for direct loading onto the BioPlex 2200 system (Bio-Rad). A result of <10 U/mL and ≥10 U/mL was interpreted as negative and positive respectively. DBS punching and elution protocols were validated using an earlier DBS panel [9 ] and took into consideration minimal input volumes for the BioPlex 2200 system.
5
Avioq HIV-1 Microelisa System Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Avioq HIV-1 Microelisa System (Avioq Inc., Research Triangle Park, NC) was used as the reference test for the qualitative detection of antibodies to HIV. DBS samples were punched (1 X 6 mm) using a pneumatic Dried Blood Spot Punch System (Analytical Sales and Services Inc., Flanders, NJ) into 2 mL 96-well polypropylene plates (ThermoFisher Scientific, Ottawa, Canada) and processed according to instructions provided by the manufacturer. Cutoff values (COV) were calculated as follows: COV = NCX + 0.270 where NCX represents the mean of the Negative Calibrator (Avioq Inc.) values. A result < COV and ≥ COV was interpreted as negative and positive respectively.
6
Dried Blood Spot HIV Antibody Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The VIDAS HIV Duo Quick (bioMérieux) was used as the index test for the qualitative detection of antibodies to HIV. DBS specimens were punched (1 X 6 mm) using a pneumatic Dried Blood Spot Punch System (Analytical Sales and Services Inc.) into 2 mL 96-well polypropylene plates (ThermoFisher Scientific). DBS punches were eluted in 200 µL of DPBS pH 7.4 containing 0.5% BSA and 0.05% Tween 20 at room temperature for 1 h with agitation (1000 rpm). The elution buffer is based on some of our previous work9 (link). The punching and elution protocol was carried out according to the Avioq HIV-1 Microelisa System package insert and took into consideration the minimal input volume for the VIDAS HIV Duo Quick test. Afterwards, all remaining DBS eluate was transferred into HIV6 Strips (bioMérieux) and processed by an automated VIDAS benchtop immunoanalyzer (bioMérieux). A test value < 0.25 and ≥ 0.25 was interpreted as negative and positive respectively unless stated otherwise. Test values (TV) were calculated as follows: TV = relative fluorescent value/background fluorescence value.
7
Serological Assay for SARS-CoV-2 Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DBS specimens were tested using an in-house CLIA at the Lunenfeld-Tanenbaum Research Institute (Mount Sinai Hospital, Sinai Health, Toronto, Canada) according to laboratory developed protocols validated during earlier work [23 ]. DBS specimens were punched (1 × 6 mm) manually or with a semi-automated BSD6000 Ascent puncher (BSD Robotics) into 2 mL 96-well polypropylene plates (ThermoFisher Scientific). DBS punches were eluted in 160 μL of PBS containing 0.1% Tween 20 and 1% Triton X-100 for a minimum of 4 h with agitation (150 rpm). Afterwards, DBS eluates were centrifuged at 1,000 x g for 30 s, transferred to new 2 mL 96-well plates, and diluted in 1.3% Blocker BLOTTO buffer (ThermoFisher Scientific) using a 1:4 ratio. Results were interpreted as follows. Spike: a relative ratio (RR) < 0.46 and ≥0.46 was interpreted as negative and positive respectively; RBD: a RR < 0.27 and ≥0.27 was interpreted as negative and positive respectively; nucleocapsid: a RR < 0.74 and ≥0.74 was interpreted as negative and positive respectively. Cutoffs represent three standard deviations from the mean of the log10 distribution of the RR from the negative controls for each antigen [23 ].
8
SARS-CoV-2 Antibody Detection in DBS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DBS specimens were tested using the Elecsys Anti-SARS-CoV-2 (nucleocapsid) and Elecsys Anti-SARS-CoV-2 S (RBD) assays (Roche Diagnostics, Basel, Switzerland) at the National Microbiology Laboratory (NML; Public Health Agency of Canada, Winnipeg, Canada) according to the manufacturer’s instructions. DBS samples were punched (4 × 6 mm) using a semi-automated BSD600 Ascent puncher (BSD Robotics) into 2 mL 96-well polypropylene plates (ThermoFisher Scientific). DBS punches were eluted in 370 μL of DPBS containing 0.5% BSA and 0.05% Tween 20 overnight at 4 °C with agitation (400 RPM). Afterwards, plates were incubated at room temperature for 30 min with agitation (400 RPM) and 250 μL of DBS eluate was transferred into 2 mL microtubes for direct loading onto a cobas e 411 analyser (Roche Diagnostics). A result of <0.8 U/mL and ≥0.8 U/mL was interpreted as negative and positive respectively with the Elecsys Anti-SARS-CoV-2 S. A cutoff index <1.0 and ≥1.0 was interpreted as negative and positive respectively with the Elecsys Anti-SARS-CoV-2 assay. DBS punching and elution protocols were validated using an earlier DBS panel [9 ].
9
Hepatitis C Antibody Detection from Dried Blood Spots
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Ortho HCV v3.0 ELISA Test System (Ortho-Clinical Diagnostics Inc., Raritan, NJ) was used as the reference test for the qualitative detection of antibodies to HCV. DBS specimens were punched (1 X 6 mm) using a pneumatic Dried Blood Spot Punch System (Analytical Sales and Services Inc.) into 2 mL 96-well polypropylene plates (ThermoFisher Scientific). DBS punches were eluted in 200 µL of Specimen Diluent (Ortho-Clinical Diagnostics Inc.) overnight at 4 °C without agitation. Afterwards, plates were gently agitated (400 rpm) at room temperature for 30 min and 150 µL of DBS eluate was transferred into HCV Encoded Antigen Coated Microwell Plates (Ortho-Clinical Diagnostics Inc.) containing 50 µL of Specimen Diluent (Ortho-Clinical Diagnostics Inc.) and processed according to instructions provided by the manufacturer. COV were calculated as follows: COV = NCX + 0.600 where NCX represents the mean of the Negative Control (Ortho-Clinical Diagnostics Inc.) values. The upper and lower limit of the grey zone (UGZ and LGZ respectively) were calculated as follows: UGZ = COV X 1.5 and LGZ = COV X 0.9. A result < LGZ, ≥ LGZ to < UGZ, and ≥ UGZ was interpreted as negative, indeterminate, and positive respectively.
10
Quantifying Biofilm Formation in Salmonella
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Typhimurium (1.0 × 106 CFU/mL) was incubated with or without GXYX crude lipopeptides at 37 °C for 48 h in a sterile 96-well polypropylene plate (Nunc, Roskilde, Denmark). The unbound cells were then washed away with sterilized PBS. Subsequently, the adhered bacteria were fixed using methanol and stained with 0.5% (m/v) crystal violet for 1 min. Thirty-three percent acetic acid was used as the decoloring solution. The absorbance at 590 nm was measured to evaluate biofilm growth [12 (link)].
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!