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SARS-CoV-2 Variants Total IgG 6-plex Panel (Flow Cytometry Multiplex Bead Assay)

For research use only.

产品描述(Product details)

Assay Type FACS
Analyte Wild-type
Alpha B.1.1.7
Beta B.1.351
Gamma P.1
Delta B.1.617.2
Omicron BA.1
Format 96T
Reactivity Human
Regulatory Status RUO
Sensitivity 16.56 ng/mL
18.33 ng/mL
14.64 ng/mL
16.23 ng/mL
15.96 ng/mL
12.81 ng/m
Standard Curve Range 32.5~1040 ng/mL
Assay Time 1 hr 50 min
Suitable Sample Type human serum, human plasma
Sample volume 3 μL

组分(Materials provided)

ID Components Size
FCM01-C01 Calibrator 20μg
FCM01-C02 RBD 6-plex Beads 200μL
FCM01-C03 Assay Buffer 40mL×2
FCM01-C04 5 × Wash Buffer 10mL
FCM01-C05 Detection Antibody 200μL
FCM01-C06 5 × PBS Buffer 10mL
FCM01-C07 APC Beads 500μL
FCM01-C08 PE Beads 500μL
FCM01-C09 96-Well V-bottom Plate 1plate
FCM01-C10 96-Well Sealing Film 2pieces

背景(Background)

Since December 2019, the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its associated disease, COVID-19, has caused a devastating pandemic worldwide.As the virus spreads globally, the continuous emergence of new mutant strains escalated the challenge on humans.To facilitate the mutant-related research, drug trials and vaccine development, a multiplex serological test for assessing antibody responses to mutant strains is in urgent need.

应用说明(Application)

The kit is developed for qualitative detection of Anti-SARS-CoV-2 Variants (Alpha, Beta, Gamma, Delta, Omicron) total IgG antibody in human serum.

It is for research use only.

存储(Storage)

1. Unopened kit should be stored at 2℃-8℃ upon receiving.
2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.
3. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

原理(Assay Principles)

SARS-CoV-2 Variants Total IgG 6-plex Panel is a multiplex bead-based indirect immunofluorescence assay, utilizing flow cytometry for data acquisition and classification. Six SARS-CoV-2 Spike RBD protein fragments (WT / Alpha / Beta / Gamma / Delta / Omicron) were conjugated to 6 fluorochrome-encoded microspheres respectively. The Anti-SARS-CoV-2 IgG Antibody in blood specimens was bound to the RBD encapsulated 6-plex beads, and then detected by PE-conjugated anti-human IgG detection antibody, forming bead-analyte-detection antibody sandwiches. The intensity of PE fluorescence was assessed by flow cytometry at wavelength of 575 nm approximately, in proportion to the Anti-SARS-CoV-2 IgG titer in blood specimens. The intensity of APC fluorescence was applied to classify bead populations, at wavelength of 670 nm approximately.

*Optional: If there is vacuum manifold for 96-Well plates, 96-Well Filter Plate (ACROBiosystems, cat#FCM-B01M) is prepared.
 

活性(Bioactivity)-FACS

Biotinylated Human FACS

A representative datapoint showing single bead population in P1 gate (left), SARS-CoV-2 variant immobilized beads classified by APC channel (middle, P2 ~ P7 gate), and measured fluorescence signal for each population in PE channel (right). Data was generated on BD FACSLyricTM Flow Cytometer, equipped with blue laser (488 nm) and red laser (640 nm).

Protocol

Spike RBD FACS

A 4-parameter logistic model fitting curve was plotted on SARS-CoV-2 Variants total IgG 6-plex Panel. Immobilized SARS-CoV-2 variant RBD proteins bind with a serial dilution of potent and broad antibody calibrator, ranging from 16.5 ng/ml to 4160 ng/ml (QC tested). EC50 values indicate the concentration at which 50% of calibrator were bound.

Protocol

 

前沿进展

 
 
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