Delta rbd

Manufactured by ACROBiosystems

The Delta RBD is a recombinant protein that corresponds to the receptor-binding domain (RBD) of the SARS-CoV-2 Delta variant spike protein. It is produced using a mammalian expression system.

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Lab products found in correlation

Alpha rbd, by ACROBiosystems (2 mentions) Beta rbd, by ACROBiosystems (2 mentions) Omicron rbd, by Sino Biological (1 mentions) Protein a biosensor, by Molecular Devices (1 mentions) Omicron rbd, by ACROBiosystems (1 mentions) Octet red384 instrument, by Molecular Devices (1 mentions) Prism 9, by GraphPad (1 mentions) 1 step ultra tmb elisa substrate, by Thermo Fisher Scientific (1 mentions) Wallac 1420 workstation, by PerkinElmer (1 mentions) Victor 5 multilabel reader, by PerkinElmer (1 mentions)

3 protocols using delta rbd

SARS-CoV-2 RBD Protein Binding Assay

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RBD‐hFc protein (Prototype RBD SPD‐C5255; Alpha RBD, SPD‐C5253; Beta RBD SPD‐C5256; Delta RBD, SPD‐C525d, purchased from ACRO Biosystems; Omicron RBD, 40592‐V05H3 Sino Biological, purchased from Sino Biological) was diluted to 1 μg/ml with PBS buffer, then each concentration gradient was diluted twice; total of eight concentration gradients were set. 293T‐ACE2 cells were collected into flow tubes (2.5 × 10⁵ cells/tube). After washing once with PBS, the liquid was poured out for use. Add 100 μl of the RBD diluent in the previous step to the flow tube, respectively, and incubate at 37°C for 40 min. Add 2 ml of PBS to wash unbound proteins, and add 1 μl of Fc Tag flow antibody (BioLegend USA) to each tube for staining at 4°C for 30 min. The excess antibody was washed with PBS, and 500 μl of PBS solution was added for detection on the machine.

Yang H., Liu P., Zhang Y., Du T., Zhou Y., Lu S, & Peng X. (2022). Characteristic analysis of Omicron‐included SARS‐CoV‐2 variants of concern. MedComm, 3(2), e129.

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Binding Affinity Analysis of SARS-CoV-2 RBD Variants

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BLI experiments were run on an Octet Red 384 instrument (Fortebio). To measure the binding affinities of monoclonal antibodies, monoclonal antibodies were immobilized onto Protein A biosensors (Fortebio) and threefold serial dilutions of WT RBD, Alpha RBD (ACROBiosystems, cat. no. SPD-C52Hn), Beta RBD (ACROBiosystems, cat. no. SPD-C52Hp), Gamma RBD (ACROBiosystems, cat. no. SPD-C52Hr), Delta RBD (ACROBiosystems, cat. no. SPD-C52Hh) and Omicron RBD (ACROBiosystems, cat. no. SPD-C522e) in PBS were used as analytes. Data were then analysed using software Octet BLI Analysis 12.2 (Fortebio) with a 1:1 fitting model. For the competitive assay by BLI, SARS-CoV-2 WT RBD tagged with His (ACROBiosystems, cat. no. SPD-C52H3) was loaded on NTA biosensors, which were pre-equilibrated in the buffer for at least 1 min. The loaded biosensors were immersed with the first monoclonal antibody for 300 s, followed by addition of the second monoclonal antibody for another 300 s. Data obtained were also analysed by Octet BLI Analsis 12.2.

Wang K., Jia Z., Bao L., Wang L., Cao L., Chi H., Hu Y., Li Q., Zhou Y., Jiang Y., Zhu Q., Deng Y., Liu P., Wang N., Wang L., Liu M., Li Y., Zhu B., Fan K., Fu W., Yang P., Pei X., Cui Z., Qin L., Ge P., Wu J., Liu S., Chen Y., Huang W., Wang Q., Qin C.F., Wang Y., Qin C, & Wang X. (2022). Memory B cell repertoire from triple vaccinees against diverse SARS-CoV-2 variants. Nature, 603(7903), 919-925.

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SARS-CoV-2 RBD Protein Binding Assay

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The following His-tagged recombinant RBD proteins (SEC-MALS verified) were purchased from Acrobiosystems, including (1) original RBD (#SPD-C52H3), (2) Kappa RBD bearing L452R and E484Q (#SPD-C52Hv), and (3) Delta RBD bearing L452R and T478K (#SPD-C52Hh). Serially diluted human sera were added to 96-well microtiter plates pre-coated with 1 μg/mL of recombinant RBD protein and were incubated at room temperature for 1h. For avidity assays, the plates were washed and overlaid with 100 μL/well of 4 M urea for 15 min.⁷⁴ (link) The plates were washed and re-blocked in blocking buffer for another hour. Bound IgG was detected using peroxidase-conjugated goat anti-human IgG (H + L) (Seracare #5220-0330, 1:2000) followed by 1-Step Ultra TMB-ELISA substrate (ThermoFisher # 34028). Optical density (OD) at 450 nm was measured using a Victor V multilabel reader (PerkinElmer) equipped with Wallac 1420 Workstation (Version 3 Revision 4). Endpoint ELISA titers were calculated based on OD values > 2-fold of blank. Avidity index was calculated as the area under curve (AUC) using Prism 9.3.1 (GraphPad).

Kwon H.J., Kosikova M., Tang W., Ortega-Rodriguez U., Radvak P., Xiang R., Mercer K.E., Muskhelishvili L., Davis K., Ward J.M., Kosik I., Holly J., Kang I., Yewdell J.W., Plant E.P., Chen W.H., Shriver M.C., Barnes R.S., Pasetti M.F., Zhou B., Wentworth D.E, & Xie H. (2022). Enhanced virulence and waning vaccine-elicited antibodies account for breakthrough infections caused by SARS-CoV-2 delta and beyond. iScience, 25(12), 105507.

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