ELISA was used to determine anti-OVA or anti-SARS-CoV-2 spike IgG titers. Briefly, 96-well microtiter plates (Thermo Fisher) were coated with 2.0 µg/mL of OVA (Invivogen) or SARS-CoV-2 spike protein (RayBiotech) overnight at 4°C. Plates were washed with 0.05% Tween-20 in PBS (PBST) and blocked with 1% BSA/PBST. Mouse serum samples were two-fold serially diluted in PBST, added to the blocked plates, and incubated at 37 °C for 1 h. Following incubation, plates were washed with PBST and incubated with horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG1 (Southern Biotech, cat#1070-05, 1:5000) or goat anti-mouse IgG2c (Southern Biotech, cat# PA1-29288, 1:4000) for 1 h. Plates were washed with PBST and TMB substrate (BD Bioscience) was added. Reactions were stopped with 50 µl 2 N H2SO4. Plates were read at OD 450 nm with a SpectraMax Plus plate reader (Molecular Devices). The antibody titer is defined as the dilution in which absorbance is more than 2.1 times of the blank wells.
Sars cov 2 spike protein
Manufactured by RayBiotech
The SARS-CoV-2 spike protein is a structural component of the SARS-CoV-2 virus. It is responsible for the attachment and entry of the virus into host cells. The spike protein is an important target for various COVID-19 diagnostic and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ovalbumin (ova), by InvivoGen (2 mentions)
96 well microtiter plate, by Thermo Fisher Scientific (2 mentions)
Tmb substrate, by BD (2 mentions)
Complete freund adjuvant (cfa), by InvivoGen (2 mentions)
B16 f10 tumor neoantigen peptides, by GenScript (2 mentions)
Endofit ovalbumin, by InvivoGen (2 mentions)
Addavax, by InvivoGen (2 mentions)
Alexa fluor 647 conjugated ova, by Thermo Fisher Scientific (2 mentions)
Horseradish peroxidase hrp conjugated goat anti mouse igg1, by Southern Biotech (1 mentions)
Goat anti mouse igg2c, by Southern Biotech (1 mentions)
Spectramax plus, by Molecular Devices (1 mentions)
Celltrace violet, by Thermo Fisher Scientific (1 mentions)
Spectramax plus plate reader, by Molecular Devices (1 mentions)
4 protocols using sars cov 2 spike protein
1
Quantifying Anti-OVA and Anti-SARS-CoV-2 IgG Titers
2
SARS-CoV-2 Spike Protein Vaccine Evaluation
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EndoFit Ovalbumin, CFA, AddaVax, and poly(I:C) were purchased from InvivoGen. The SARS-CoV-2 spike protein was purchased from RayBiotech. Alexa FluorTM 647 conjugated OVA and CellTrace Violet were purchased from Thermo Fisher. B16-F10 tumor neoantigen peptides were synthesized by GenScript (Piscataway, NJ). The sequences are as follows: M27: REGVELCPGNKYEMRRHGTTHSLVIHD; M30: PSKPSFQEFVDWENVSPELNSTDQPFL; M48: SHCHWNDLAVIPAGVVHNWDFEPRKVS.
3
ELISA-based Antibody Titer Determination
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ELISA was used to determine anti-OVA or anti-SARS-CoV-2 spike IgG titers. Briefly, 96-well microtiter plates (Thermo Fisher) were coated with 2.0 µg/mL of OVA (Invivogen) or SARS-CoV-2 spike protein (RayBiotech) overnight at 4˚C. Plates were washed with 0.05% Tween-20 in PBS (PBST) and blocked with 1% BSA/PBS-T. Mouse serum samples were two-fold serially diluted in PBST, added to the blocked plates, and incubated at 37°C for 1 h. Following incubation, plates were washed with PBS-T and incubated with horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG1 or goat anti-mouse IgG2c (Southern Biotech) for 1 h. Plates were washed with PBS-T and TMB substrate (BD Bioscience) was added. Reactions were stopped with 50 µl 2N H2SO4. Plates were read at OD 450 nm with a SpectraMax Plus plate reader (Molecular Devices).
The antibody titer is defined as the dilution in which absorbance is more than 2.1 times of the blank wells.
The antibody titer is defined as the dilution in which absorbance is more than 2.1 times of the blank wells.
4
Neoantigen-Based Tumor Vaccination Protocol
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EndoFit Ovalbumin, CFA, AddaVax, and poly(I:C) were purchased from InvivoGen. The SARS-CoV-2 spike protein was purchased from RayBiotech. Alexa FluorTM 647 conjugated OVA was purchased from Thermo Fisher. B16-F10 tumor neoantigen peptides were synthesized by GenScript (Piscataway, NJ). The sequences are as follows: M27: REGVELCPGNKYEMRRHGTTHSLVIHD; M30: PSKPSFQEFVDWENVSPELNSTDQPFL; M48: SHCHWNDLAVIPAGVVHNWDFEPRKVS using the mutation information described 4 (link) .
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