The largest database of trusted experimental protocols

35 protocols using m1000 pro plate reader

1

Curcumin-Loaded Polymer Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols
Curcumin was loaded to SMA and FA-DABA-SMA polymers through physical entrapment. Briefly, excess curcumin was added to polymer stock solutions at 54 μm concentration. The solution was allowed to settle and was centrifuged at 4,000 rpm (2500× g) for 7 minutes to remove the remaining curcumin from the solution. Curcumin-loaded polymers were then diluted to four different concentrations of 0.3, 1, 3, and 10 μm. The curcumin-loaded SMA was characterized using Tecan M1000 Pro plate reader (Tecan Group Ltd, Männedorf, Switzerland) in 96-well microplates.
+ Open protocol
+ Expand
2

Quantification of Protochlorophyllides in Arabidopsis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protochlorophyllides (Pchlides) were extracted and measured using published methods (Kolossov and Rebeiz, 2003 (link)) with modifications. Around 100 mg of etiolated Arabidopsis seedlings (7-d-old) were harvested under dim-green safe light, frozen and ground to fine powder. Two milliliters of 80% ice-cold acetone was added to each sample and the mixture was briefly homogenized. After centrifugation at 18,000 g for 10 min at 1 ⁰C, supernatant was split to 2 × 1 mL for Pchlides and protein extraction. Fully esterified tetrapyrroles were extracted from the acetone extracts with equal volume followed by 1/3 vol of hexane. Pchlides remained in the hexane-extracted acetone residue were used for fluorescence measurement with a TECAN M1000PRO plate reader (Tecan Group) and net fluorescence were determined as previously described (Rebeiz et al., 1975 (link)). Protein extraction was performed using 80% acetone and 10% TCA; protein concentration was used to normalize the net fluorescence of Pchlides.
+ Open protocol
+ Expand
3

SARS-CoV-2 Spike RBD IgG Antibody ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols
ELISA plates (Thermo Fisher #3455) were coated overnight at 4°C with 2 μg/mL of SARS-CoV-2 Spike RBD (Sino Biological #40592-V08B) in bicarbonate buffer (Sigma-Aldrich #88975) and blocked by 1% BSA in PBS with 0.1% Tween 20 (PBST) for 1 hr at room temperature. Serum samples were 3-fold serially diluted (in blocking buffer), starting at 1:100, and loaded to each well (100 μL). Following incubation at 37°C for 60 minutes, plates were washed 3 times with 1× PBS. Then 100 μL of diluted anti-mouse IgG-peroxidase rabbit antibody (Sigma-Aldrich #A9044, 1:10,000 dilution) was added per well and incubated at 37°C for 30 minutes. Plates were washed 3 times following incubation, and TMB substrates were added (100 μL/well) and incubated for 15-20 minutes at room temperature. Then, the ELISA stop buffer was added and the absorbance (450/630 nm) was measured with the TECAN infinite M1000 Pro plate reader. The endpoint IgG titers were defined as the highest dilution fold of sera to yield an absorbance greater than 2.1-fold of the background values (without serum but the secondary antibody was added) as previously described.27 (link) Antibody titer below the limit of detection was determined as half the limit of detection.
+ Open protocol
+ Expand
4

Evaluating CrCl3 Cytotoxicity with MTT Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The effects of CrCl3 on cellular viability were assessed
using the MTT cell proliferation
assay.68 (link),69 (link) Briefly, 2 × 104 cells per
well were seeded onto Corning Scientific Costar 96-well polystyrene
flat bottom plates. After 24 h, the growth medium was aspirated and
replaced with new growth medium containing CrCl3. Sodium
dodecyl sulfate (0.05, 0.10, 0.15, or 0.20 mg/mL) was used as a positive
control for the assay.
After incubation for 1 h, the growth
medium was carefully aspirated and replaced with 100 μL of fresh
growth medium (absent of phenol red) and 10 μL of the MTT reagent.
After 6 h, the MTT solution was carefully aspirated and the formazan
crystals were dissolved with 50 μL of spectroscopic grade DMSO.
The absorbance at 540 nm was read by an M1000 PRO plate reader (Tecan,
Switzerland) following 1 s of shaking at 2 mm amp and 654 rpm. To
remove the background media effects, empty (blank) wells were also
treated according to the MTT protocol.
+ Open protocol
+ Expand
5

Cell Proliferation Assay via GFP Fluorescence

Check if the same lab product or an alternative is used in the 5 most similar protocols
To determine cell proliferation rate using a fluorescence plate reader, TNBC cells were stably transfected with GFP (pRetroQ‐AcGFP1‐N1). Transfected cells were selected with puromycin (BT‐20 at 1.5 μg/ml, 468 at 0.5 μg/ml, and 231 at 2 μg/ml). Cells were selected until a parallel non‐transformed plate exposed to puromycin was completely dead. The selected population was subsequently sorted by FACS to collect cells with similar levels of GFP fluorescence. For coculture experiments, fibroblast cell lines were plated at an 8:1, 4:1, 2:1, 1:1, and 1:2 ratio to cancer cells in a Greiner 96‐well plate in 100 μl of FluoroBrite media and allowed to adhere for 3 h. Following adherence of fibroblast cells, TNBC cells constitutively expressing GFP were plated at a concentration of 10,000 cells per 100 μl of FluoroBrite media and allowed to adhere overnight. Cell measurements were measured every 24 h for 96 h using a Tecan M1000 Pro Plate reader.
+ Open protocol
+ Expand
6

3D Culture and Drug Response Assay for TNBC

Check if the same lab product or an alternative is used in the 5 most similar protocols
Culturing of TNBC cells in 3D colonies was performed using the “3D on top” method developed by Bissell and colleagues (Lee et al, 2007). Briefly, a thick layer of cold Matrigel (corning cat#356235) was applied to the bottom of 96 well plates, which were subsequently heated to 37°C for 30 min to promote solidification. Cancer cells were plated at a concentration of 10,000 cells in 100 μl complete media + 2% Matrigel and were grown for 72 h to induce 3D colony formation. After 72 h of growth, the media were aspirated and media containing drug + 2% Matrigel were added to each well. At 72 h post‐drug addition, cell viability was measured by adding 100 μl of CellTiter‐Glo reagent to each well. The CellTiter‐Glo assay was performed according to the manufacturer's directions, and luminescence was read using a Tecan M1000 Pro Plate Reader.
+ Open protocol
+ Expand
7

Quantifying Cardiac Creatine Kinase Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
After cervical dislocation, blood was collected from the heart, placed into cryovials, centrifuged at 4000 rpm for 10 min at 4 °C, and serum collected. A colorimetric creatine kinase activity assay kit was used to determine CK-MM levels (Abcam, Melbourne, Vic. Australia Cat. No.: ab155901). To determine optimal diluent concentration for the assay, a standard curve was created and then samples were run in duplicate and were measured at OD 450 nm on a Tecan M1000 Pro Plate Reader in a kinetic mode, every 1 min for 40 min at 37 °C.
+ Open protocol
+ Expand
8

Spectrophotometric Assay for TPI Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
TPI activity was determined using a spectrophotometric assay as previously described.29, 38 Measurements were performed at 2‐3 different protein concentrations per tissue lysate and repeated independently three times with freshly dissected tissues from 20‐week‐old male mice. NADH loss was measured using a Tecan M1000 PRO plate reader. TPI activity was calculated at the point of maximal reaction rate and normalized to lysate‐free background activity.
+ Open protocol
+ Expand
9

Hemin-Catalyzed Oxidation of ABTS

Check if the same lab product or an alternative is used in the 5 most similar protocols
Hemin was dissolved in DMSO to obtain a solution at 1 mM concentration and stored in the dark at –20°C. ABTS2−and H2O2were freshly prepared in water at the desired concentration for use. DNA samples were folded by heating at 95°C for 5 min in the corresponding buffer and left to cool at room temperature for at least two hours. 5 μl of a fresh dilution of Hemin at 60 μM (10% DMSO) were then added to reach a final Hemin concentration of 6 μM and the solutions were left to stand at room temperature (25°C) for 30 min. 5 μl ABTS2− at 5 mM was then added to each sample to a final concentration of 500 μM, and basal absorbance was measured. To initiate the oxidation reaction, H2O2 was added to a final concentration of 50 μM, followed by quick mixing. 15 min later, the absorbance at 420 nm of the oxidized product ABTS•− was monitored by a TECAN M1000 pro plate reader (France). The final sample contained 3 μM DNA, 6 μM Hemin, 1% DMSO, 500 μM ABTS2− and 50 μM H2O2 in a total volume of 50 μl.
+ Open protocol
+ Expand
10

Membrane Interactions of P0ct Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols
For turbidimetric measurements, SUVs of 0.5 mM DOPC:DOPG (1:1) and DMPC:DMPG (1:1), both with and without supplemented 10% (w/w) cholesterol, were mixed with 0.5–10 µM P0ct in duplicate. Light scattering was recorded at 450 nm for 10 min at 25 °C using a Tecan M1000Pro plate reader. The results were analyzed after the observed optical density per time had stabilized.
SAXD experiments were performed to investigate any repetitive structures in turbid samples. 10 and 20 µM P0ct was mixed with SUVs of 1–3 mM DMPC:DMPG (1:1) in HBS at ambient temperature and exposed at 25 °C on the EMBL P12 BioSAXS beamline, DESY (Hamburg, Germany). A HBS buffer reference was subtracted from the data. Lipid samples without added P0ct were devoid of Bragg peaks. The peak positions of momentum transfer, s, in P0ct-lipid samples were used to calculate mean real-space repeat distances, d, in proteolipid structures, using the equation d=2πs,where=s=4πsinθλ.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!