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S trimer

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The S trimer is a lab equipment product provided by ACROBiosystems. It is a recombinant protein that represents the trimeric spike (S) protein of the SARS-CoV-2 virus. The core function of the S trimer is to serve as a research tool for studying the structure and interactions of the SARS-CoV-2 spike protein.

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Lab products found in correlation

Ab207203, by Abcam (2 mentions) Ab14860, by Abcam (2 mentions) S rbd, by ACROBiosystems (2 mentions) Hbs ep buffer, by Cytiva (1 mentions) Biacore 8k system, by Cytiva (1 mentions) Amine coupling kit, by Cytiva (1 mentions) Cm5 sensor chip, by Cytiva (1 mentions) Streptavidin peroxidase polymer ultrasensitive, by Merck Group (1 mentions) Sars cov 2 rbd, by Sino Biological (1 mentions) Ez link sulfo nhs biotin, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions)

Close spelling variants of this product

SARS-CoV-2 S protein trimer

4 protocols using s trimer

1

Spike Protein Binding Kinetics

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The simultaneous binding of four antibodies onto the spike protein was tested using a BIAcore 8K system (Cytiva) together with CM5 sensor chips (Cytiva). S trimer (AcroBiosystems) was diluted in pH 5.0 Acetate Buffer (Cytiva) and covalently immobilized on chips using an Amine Coupling Kit (Cytiva). After reaching a ∼ 270 RU coupling level (correspond to ​∼ ​100 RU level of Rmax), the excess antigens were washed away and the unbound sites were blocked with ethanolamine. Antibodies were diluted with HBS-EP buffer (Cytiva) to the concentration of 40 ​μg/mL. The first antibody was injected at a rate of 20 ​μL/min for 600 ​s to reach a saturated binding level, followed by injecting a second antibody into the system. The ascending RU value was monitored to detect the successive binding of four antibodies.
Tang H., Ke Y., Liao Y., Bian Y., Yuan Y., Wang Z., Yang L., Ma H., Sun T., Zhang B., Zhang X., Wu M, & Zhu J. (2022). Mutational escape prevention by combination of four neutralizing antibodies that target RBD conserved regions and stem helix. Virologica Sinica, 37(6), 860-873.
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2

Evaluating ERα Activity in MCF-7 Cells

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The ERα activity was determined by the ERα transcription factor activation assay kit (Abcam, ab207203) according to the manufacturer’s directions. Briefly, MCF-7 nuclear extracts (5 μg; Abcam, ab14860) were treated with S (0.01 to 300 nM; ACROBiosystems), S-RBD (1 to 100 nM; ACROBiosystems), S-trimer (1 to 100 nM; provided by M. Borgnia), and/or E2 (100 nM: Tocris). Extracts were added to each well coated with the ER consensus binding site (5′-GGTCACAGTGACC-3′). The wells were washed and then incubated with rabbit anti-ERα (1:2000; 1 hour at RT) and horseradish peroxidase (HRP)–conjugated secondary antibody (1:2000; 1 hour at RT) that were provided with the kit. Colorimetric reaction was measured by spectrophotometry at a wavelength of 450 nm. Values were given as percentage of the effect induced by E2.
Solis O., Beccari A.R., Iaconis D., Talarico C., Ruiz-Bedoya C.A., Nwachukwu J.C., Cimini A., Castelli V., Bertini R., Montopoli M., Cocetta V., Borocci S., Prandi I.G., Flavahan K., Bahr M., Napiorkowski A., Chillemi G., Ooka M., Yang X., Zhang S., Xia M., Zheng W., Bonaventura J., Pomper M.G., Hooper J.E., Morales M., Rosenberg A.Z., Nettles K.W., Jain S.K., Allegretti M, & Michaelides M. (2022). The SARS-CoV-2 spike protein binds and modulates estrogen receptors. Science Advances, 8(48), eadd4150.
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3

SARS-CoV-2 Antigen-Antibody Binding Assay

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For experiments involving the competitive binding of antibodies to SARS-CoV-2 RBD or S trimer, recombinant hACE2-Fc protein was first biotinylated using EZ-Link Sulfo-NHS-Biotin (ThermoFisher) as the instruction described. SARS-CoV-2 RBD (Sino Biological), S trimer (AcroBiosystems), mutated RBDs (Sino Biological), and mutated S trimers (AcroBiosystems) were coated onto High Binding ELISA 96-Well Plate (BEAVER). In order to obtain an optimized hACE2-Fc concentration for this experiment, the concentration-dependent binding of biotinylated hACE2-Fc to coated SARS-CoV-2 antigens was measured by performing a conventional receptor-binding ELISA. The 80% maximal effective concentration (EC80) of biotinylated hACE2-Fc was calculated by the four-parameter nonlinear fitting. Antibodies were serially diluted in 1% BSA (Sigma) and added 50 μL into the antigen coated plates. Biotinylated hACE2-Fc at EC80 concentration was subsequently pipetted into. After incubation at 37 °C for 1 h, plates were washed four times with PBST and incubated with 100 μL of 1:2000 diluted Ultrasensitive Streptavidin-Peroxidase Polymer (Sigma). After further washing, 100 μL TMB was added, followed by detection of the bound hACE2 in the microplate reader. Four-parameter nonlinear regression fitting in GraphPad Prism Version 9.0.0 was applied for result analysis.
Ma H., Guo Y., Tang H., Tseng C.T., Wang L., Zong H., Wang Z., He Y., Chang Y., Wang S., Huang H., Ke Y., Yuan Y., Wu M., Zhang Y., Drelich A., Kempaiah K.R., Peng B.H., Wang A., Yang K., Yin H., Liu J., Yue Y., Xu W., Zhu S., Ji T., Zhang X., Wang Z., Li G., Liu G., Song J., Mu L., Xiang Z., Song Z., Chen H., Bian Y., Zhang B., Chen H., Zhang J., Liao Y., Zhang L., Yang L., Chen Y., Gilly J., Xiao X., Han L., Jiang H., Xie Y., Zhou Q, & Zhu J. (2022). Broad ultra-potent neutralization of SARS-CoV-2 variants by monoclonal antibodies specific to the tip of RBD. Cell Discovery, 8, 16.
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4

Assay of ERα Transcriptional Activity

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The ERα activity was determined by the ERα transcription factor activation assay kit (ab207203, Abcam) according to manufacturer’s directions. Briefly, MCF-7 nuclear extracts (5 μg; ab14860, Abcam) were treated with either S (0.01–300 nM; Acro Biosystems) S-RBD (1–100 nM; Acro Biosystems), S-trimer (1–100 nM; provided by Dr. Borgnia) and/or E2 (100 nM: Tocris). Extracts were added to each well coated with the ER consensus binding site (5’ – GGTCACAGTGACC – 3’). The wells were washed and then incubated with rabbit anti-ERα (1:2000, 1 h, RT) and horseradish-conjugated secondary antibody (1:2000, 1 h, RT) that were provided with the kit. Colorimetric reaction was measured by spectrophotometry at a wavelength of 450 nm.
Solis O., Beccari A.R., Iaconis D., Talarico C., Ruiz-Bedoya C.A., Nwachukwu J.C., Cimini A., Castelli V., Bertini R., Montopoli M., Cocetta V., Borocci S., Prandi I.G., Flavahan K., Bahr M., Napiorkowski A., Chillemi G., Ooka M., Yang X., Zhang S., Xia M., Zheng W., Bonaventura J., Pomper M.G., Hooper J.E., Morales M., Rosenberg A.Z., Nettles K.W., Jain S.K., Allegretti M, & Michaelides M. (2022). The SARS-CoV-2 spike protein binds and modulates estrogen receptors. bioRxiv.
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