Biacore t200 evaluation software 3

Manufactured by Cytiva

The Biacore T200 evaluation software 3.0 is a data analysis tool that enables users to process and analyze data from the Biacore T200 biosensor instrument. It provides functionalities for data import, fitting, and visualization to support the evaluation of biomolecular interactions.

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Lab products found in correlation

Biacore t200, by Cytiva (5 mentions) Human type 4 placental collagen, by Merck Group (1 mentions) T200 instrument, by Cytiva (1 mentions) Amine coupling kit, by Cytiva (1 mentions) Biacore t200 system, by Cytiva (1 mentions)

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Biacore T200 (3299 citations) T200 Evaluation Software (49 citations) Biacore T200 Evaluation Software (36 citations) Biacore Evaluation Software (20 citations) T200 Evaluation Software 3.1 (15 citations) Evaluation software (13 citations)

5 protocols using biacore t200 evaluation software 3

SPR Analysis of Stapled Peptide Binding

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The SPR assay protocol was based on previously reported conditions, [28 (link)] with the following changes: SPR analysis was performed on a BiacoreT200; ERα ligand binding domain construct contained amino acids 299–554, including N-terminal 6His-tag; final ERα surface density was ~9500 RU; stapled peptide solutions at a series of increasing concentrations were applied to flow cells at a 30 μL/min flow rate using a contact time of 60 s and a dissociation time of 120 s; K_D values were determined by fitting reference subtracted data to a steady-state affinity equation embedded in the Biacore T200 evaluation software 3.0; and kinetic fittings were done using the two state reaction binding equation embedded in the Biacore T200 evaluation software 3.0 (Figure S8).

Speltz T.E., Danes J.M., Stender J.D., Frasor J.M, & Moore T.W. (2018). A cell-permeable stapled peptide inhibitor of the estrogen receptor/coactivator interaction. ACS chemical biology, 13(3), 676-684.

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Quantifying SARS-CoV-2 Spike Antibody Affinity

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BIAcore T200 surface plasmon resonance (SPR; Cytiva) was used to assess anti-S2 antibody affinities for the S antigen. SARS-CoV-2 spike (D614G)-ECD and SARS-CoV spike-ECD were immobilized on CM5 sensor chips by EDC/NHS activation. Antibodies were tested at six dilutions (12.5–400 nM or 6.25–100 nM) in HBS-EP+ buffer (0.01 M HEPES pH 7.4, 0.15 M NaCl, 3 mM EDTA and 0.05% Surfactant P20). Measurements were made at 30 μL/min, 25°C, in HBS-EP+ buffer. Antibody affinity was analyzed with BIAcore T200 Evaluation software 3.1 using a 1:1 (Langmuir) binding model.

Heo C.K., Lim W.H., Yang J., Son S., Kim S.J., Kim D.J., Poo H, & Cho E.W. (2023). Novel S2 subunit-specific antibody with broad neutralizing activity against SARS-CoV-2 variants of concern. Frontiers in Immunology, 14, 1307693.

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Quantifying rFIX Binding to Collagen IV

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Binding of rFIX molecules to collagen IV was measured by surface plasmon resonance (SPR), using Biacore T-200 (Cytiva Europe GmbH, Germany) and HBS-N (10 mM HEPES, 150 mM NaCl, pH 7.4) as a sample and running buffer. The analysis was performed at 25°C. Human-type IV placental collagen (Sigma, Germany) was immobilized on an active flow cell of a sensor series CM5 chip using an amine coupling chemistry kit (Cytiva) to approximately 3,300 RU. Reference flow cell was treated as blank immobilization. The immobilized collagen IV was washed four times with 10 mM HCl and once with HBS-N before injection of rFIX. Each analyte (human rFIX or human fibronectin) was injected for 180 seconds over reference and active flow cells followed by 180 seconds of dissociation at 30 µL/min. Each molecule was analyzed four times. Data were analyzed with Biacore T200 Evaluation Software 3.1.

Machado S.K., Peil H., Kraushaar T., Claar P., Mischnik M., Lind H., Herzog E., Bacher M., Nolte M.W., Bielohuby M., Pestel S, & Ponnuswamy P. (2023). Modulation of Extravascular Binding of Recombinant Factor IX Impacts the Duration of Efficacy in Mouse Models. Thrombosis and Haemostasis, 123(8), 751-762.

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SPR Analysis of ClpC1-Small Molecule Binding

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Each ClpC1 enzyme was immobilized on a CM5 sensor chip using standard amine-coupling at 25 °C with running buffer PBS-P (20 mM Phosphate, 137 mM NaCl, 27 mM KCl, 0.05% surfactant P-20, pH 7.4) using a Biacore T200 instrument. Unmodified blank surface was used on flow channel 1 as a control. ClpC1 enzymes were diluted with 10 mM sodium acetate (pH 4.0 or pH 5.0) to 50 μg/mL and immobilized to flow channels 2, 3, and 4 after sensor surface activation with EDC/NHS, followed by ethanolamine blocking on unoccupied surface area. Compound solutions with a series of increasing concentrations (0–5 μM at 2-fold dilution) were applied to all four channels in SPR binding buffer (PBS-P, 0.5 mM TCEP, 2% DMSO) at a 30 μL/min flow rate at 25 °C. For competition experiments, each titration of RUFI also contained 10 μM ECU and vice versa. Sensorgrams were analyzed using the Biacore T200 evaluation software 3.0, and data were double-referenced with reference channel and zero concentration of compound response unit values. Kinetic rate constants (k_a and k_d) were determined by fitting the double-reference data globally to the 1:1 Langmuir model embedded in the Biacore T200 evaluation software (v3.0). K_D values were then calculated from the two rate constants (K_D = k_d/k_a).

Wolf N.M., Lee H., Choules M.P., Pauli G.F., Phansalkar R., Anderson J.R., Gao W., Ren J., Santarsiero B.D., Lee H., Cheng J., Jin Y.Y., Ho N.A., Duc N.M., Suh J.W., Abad-Zapatero C, & Cho S. (2019). High-Resolution Structure of ClpC1-Rufomycin and Ligand Binding Studies Provide a Framework to Design and Optimize Anti-Tuberculosis Leads. ACS infectious diseases, 5(6), 829-840.

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SPR Analysis of Beta-Catenin Interactions

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SPR analysis was performed on a Biacore T200 system (Cytiva). HEPES-buffered saline containing 0.05% Tween-20 and 5% DMSO was used as running buffer. By using a series S CM5 sensor chip and the Amine-coupling Kit (Cytiva) as per manufacturer's instructions, purified beta-catenin was amine-coupled to flow cells (FC) #2 and #4 at 7800 and 10779 RU, respectively. Analyte samples, iCRT3 and GB1874, diluted in running buffer were then injected over all four FCs so that FC(2-1) and FC(4-3) referenced data were collected at 25 degrees Celcius. Concentration-dependent response was observed although the sensorgrams suggest a degree of sample aggregation in the running buffer. Biacore T200 Evaluation Software 3.0 was used for steady-state affinity analysis with the assumption of one-to-one binding model. For each set of sensorgrams, a time point at which steady state signal is attained is chosen, the response levels plotted against concentration, and the KD values derived.

Low J.L., Du W., Gocha T., Oguz G., Zhang X., Chen M.W., Masirevic S., Yim D.G., Tan I.B., Ramasamy A., Fan H, & DasGupta R. (2021). Molecular docking-aided identification of small molecule inhibitors targeting β-catenin-TCF4 interaction. iScience, 24(6), 102544.

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