Product Manual

SARS-CoV-2 Spike Protein S1 ELISA Kit
Catalog Number
VPK-5155

96 assays

VPK-5155-5

5 x 96 assays

FOR RESEARCH USE ONLY
Not for use in diagnostic procedures

Introduction
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel form of coronavirus. It was
first identified in three people related to the cluster of acute respiratory illness cases in Wuhan, China.
SARS-CoV-2 can have effects on the sinuses, nose, and throat (upper respiratory tract) as well as thelungs and windpipe (lower respiratory tract). The organ most affected by this virus is the lungs:SARS-CoV-2 enters host cells by binding to the enzyme angiotensin-converting enzyme 2 (ACE2),
which is abundant in type II alveolar cells located in the lungs. The virus binds to and enters humancells by using its surface spike glycoprotein (spike protein) to bind ACE2 on the host cell surface. As
the disease develops in the lungs, respiratory failure can occur and death may result. SARS-CoV-2
may also be responsible for respiratory failure by invading the brain stem since similar coronaviruseshave been found to enter the central nervous system (CNS). Although SARS-CoV-2 has been detectedin cerebrospinal fluid of the deceased, the exact mechanism of CNS invasion remains unknown. Thevirus can also enter cells in the gastrointestinal organs because ACE2 is expressed in the glandular
cells of gastric, duodenal and rectal epithelium as well as endothelial cells of the small intestine.
Finally, SARS-CoV-2 can cause myocardial injury and damage to the cardiovascular system.
SARS-CoV-2 is closely related to the original virus known as SARS-CoV. The protein makeup ofSARS-CoV-2 includes membrane glycoprotein (M), envelope protein (E), spike protein (S), and thenucleocapsid protein (N). The spike protein is a glycoprotein that is split into two parts (S1 and S2).
The spike protein S1 subunit catalyzes attachment to the mammalian cell surface while the S2 subunit
promotes fusion with the target membrane.
Cell Biolabs’ SARS-CoV-2 Spike Protein S1 ELISA Kit is an enzyme immunoassay developed for thedetection and quantitation of SARS-CoV-2 Spike Protein (S1 subunit). The kit has a detection
sensitivity limit of 31.3 ng/mL spike protein. Each kit provides sufficient reagents to perform up to 96
assays including standard curve and unknown samples.

Related Products
1. VPK-5145: SARS-CoV-2 Nucleocapsid ELISA Kit
2. VPK-5154: West Nile Virus Envelope Protein ELISA Kit
3. VPK-107: QuickTiter™ Lentivirus Titer Kit (Lentivirus-associated p24 ELISA)
4. VPK-109: QuickTiter™ Adenovirus Titer Immunoassay Kit
5. VPK-112: QuickTiter™ Lentivirus Quantitation Kit6. VPK-120: QuickTiter™ Retrovirus Quantitation Kit
7. VPK-140: ViraBind™ AAV Purification Kit
8. VPK-145: QuickTiter™ AAV Quantitation Kit
9. VPK-5112: PureVirus™ Adenovirus Purification Kit

2Kit Components
Box 1 (shipped at room temperature)
1. Anti-SARS-Cov-2 Spike S1 Ab Coated Plate (Part No. 51551B): One 96-well strip plate (8 x12).
2. Biotinylated Anti-Spike Protein S1 Ab (1000X) (Part No. 51552C): One 10 µL vial.
3. Streptavidin-Enzyme Conjugate (Part No. 310803): One 20 µL vial.
4. Assay Diluent (Part No. 310804): One 50 mL bottle.
5. Triton X-100 Solution (Part No. 310805): One 15 mL bottle containing 5% Triton X-100 inTBS.
6. 10X Wash Buffer (Part No. 310806): One 100 mL bottle.
7. Substrate Solution (Part No. 310807): One 12 mL amber bottle.
8. Stop Solution (Part. No. 310808): One 12 mL bottle.
Box 2 (shipped on blue ice packs)
1. SARS-CoV-2 Spike S1 Standard (Part No. 51553D): One 25 µL vial of 200 µg/mL
recombinant SARS-CoV-2 Spike S1 Protein.

Materials Not Supplied
1. SARS-CoV-2 Samples
2. 10 µL to 1000 µL adjustable single channel micropipettes with disposable tips
3. 50 µL to 300 µL adjustable multichannel micropipette with disposable tips
4. Multichannel micropipette reservoir
5. Microplate reader capable of reading at a wavelength of 450 nm

StorageUpon receipt, store the SARS-CoV-2 Spike S1 Standard at -80ºC and avoid multiple freeze/thawcycles. Store the Biotinylated Anti-SARS-CoV-2 Spike S1 Antibody at -20ºC or -80ºC. Store all othercomponents at 4ºC.

Preparation of Reagents•1X Wash Buffer: Dilute the 10X Wash Buffer to 1X with deionized water. Stir to homogeneity.

•Biotinylated Anti-SARS-CoV-2 Antibody and Streptavidin-Enzyme Conjugate: Immediately
before use dilute the Biotinylated Anti-SARS-CoV-2 Antibody and the Streptavidin-Enzyme
Conjugate 1:1000 with Assay Diluent. Do not store diluted solutions.

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Preparation of Standard Curve
1. Prepare a dilution series of Spike S1 standards in the concentration range of 0 to 2000 ng/mL into
Assay Diluent (Table 1).
StandardTubes
12345678200 µg/mL Spike S1Standard (μL)
5250 of Tube #1
250 of Tube #2250 of Tube #3
250 of Tube #4
250 of Tube #5
250 of Tube #60

Assay Diluent (µL)
495
250
250
250
250250250
250

Spike S1 (ng/mL)
2000
1000
500
250
125
62.5
31.3
0

Table 1. Preparation of Spike S1 Standards
2. Transfer 225μL of each dilution to a microcentrifuge tube containing 25 μL of Triton X-100Solution. Perform the assay as described in Assay Instructions.

Preparation and Inactivation of Samples
1.

(Optional) Dilute viral supernatant in culture medium as needed. For unknown samples werecommend several dilutions for each sample. Include culture medium as a negative control.

2. Transfer 225 µL of each sample to a microcentrifuge tube containing 25 µL of Triton X-100
Solution. Vortex well.
3. Incubate 30 minutes at room temperature.

Assay Protocol
1. Add 100 µL of Spike S1 unknown sample, standard, or blank to the Anti-SARS-CoV-2 Spike S1
Antibody Coated Plate. Each Spike S1 unknown sample, standard and blank should be assayed induplicate.
2. Incubate at room temperature for 1 hour on an orbital shaker.
3. Wash microwell strips 3 times with 250 µL 1X Wash Buffer per well with thorough aspiration
between each wash. After the last wash, empty wells and tap microwell strips on absorbent pad or
paper towel to remove excess 1X Wash Buffer.
4. Add 100 µL of the diluted Biotinylated Anti-Spike S1 antibody to each well. Incubate at room
temperature for 1 hour on an orbital shaker.
5. Wash the strip wells 3 times according to step 3 above.
6. Add 100 µL of the diluted Streptavidin-Enzyme Conjugate to each well.
temperature for 1 hour on an orbital shaker.
4Incubate at room

7. Wash the strip wells 3 times according to step 3 above. Proceed immediately to the next step.
8. Warm Substrate Solution to room temperature. Add 100 L of Substrate Solution to each well,including the blank wells. Incubate at room temperature on an orbital shaker. Actual incubation
time may vary from 2-30 minutes.
Note: Watch plate carefully; if color changes rapidly, the reaction may need to be stopped sooner
to prevent saturation.
9. Stop the enzyme reaction by adding 100 µL of Stop Solution into each well, including the blankwells. Results should be read immediately (color will fade over time).
10. Read absorbance of each microwell on a spectrophotometer using 450 nm as the primary wavelength.

Example of Results
The following figures demonstrate typical results with the SARS-CoV-2 Spike S1 ELISA Kit. One
should use the data below for reference only. This data should not be used to interpret actual results.

Figure 1: SARS-CoV-2 Spike Protein S1 ELISA Kit Standard Curve.

5References
1. Andersen KG, Rambaut A, Lipkin WI, Holmes EC, and Garry RF (2020). Nature Med. 26: 450–452.
2. Verdecchia P, Cavallini C, Spanevello A, and Angeli F (2020). Euro. J. Int. Med. 76: 14–20.
3. Letko M, Marzi A, and Munster V (2020). Nature Microbiol. 5: 562–569.
4. Li YC, Bai WZ, and Hashikawa T (2020). J. Med. Vir. 92: 552–555.
5. Gu J, Han B, and Wang J (2020). Gastroenterology. 158: 1518–1519.
6. Zheng YY, Ma YT, Zhang JY, and Xie X (2020). Nature Rev. Cardiol. 17: 259–260.
7. Du L, Zhao G, Lin Y, Chan C, He Y, Jiang S, Wu C, Jin D-Y, Yuen K-Y, Zhou Y, Zheng B-J(2008) Vaccine 26: 1644-1651.
8. V'kovski P, Kratzel A, Steiner S, Stalder H, and Thiel V (2021). Nature Rev. Microbiol. 19: 155–170.
9. Wu C, Liu Y, Yang Y, Zhang P, Zhong W, Wang Y, Wang Q, Xu Y, Li M, Li X, Zheng M, Chen
L, and Li H (2020). Acta Pharm. Sinica. B. 10: 766–788

Warranty
These products are warranted to perform as described in their labeling and in Cell Biolabs literature when used inaccordance with their instructions. THERE ARE NO WARRANTIES THAT EXTEND BEYOND THIS EXPRESSEDWARRANTY AND CELL BIOLABS DISCLAIMS ANY IMPLIED WARRANTY OF MERCHANTABILITY OR
WARRANTY OF FITNESS FOR PARTICULAR PURPOSE. CELL BIOLABS’ sole obligation and purchaser’s
exclusive remedy for breach of this warranty shall be, at the option of CELL BIOLABS, to repair or replace the products.
In no event shall CELL BIOLABS be liable for any proximate, incidental or consequential damages in connection with theproducts.

Contact Information
Cell Biolabs, Inc.
5628 Copley Drive
San Diego, CA 92111
Worldwide: +1 858-271-6500USA Toll-Free: 1-888-CBL-0505
E-mail: tech@cellbiolabs.com
www.cellbiolabs.com©2021-2024: Cell Biolabs, Inc. - All rights reserved. No part of these works may be reproduced in any form without
permissions in writing.

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