膜杰作
Star Staining
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 B.1.1.529 |
| Format | 96T |
| Reactivity | Human |
| Regulatory Status | RUO |
| Sensitivity |
38.78 ng/mL 42.79 ng/mL 35.7 ng/mL 39.72 ng/mL 35.74 ng/mL 54.74 ng/mL |
| Standard Curve Range | 130~16640 ng/mL |
| Assay Time | 1 hr 34 min |
| Suitable Sample Type | human serum, human plasma |
| Sample volume | 5 μL |
组分（Materials Provided）
| ID | Components | Size |
| FCM04-C01 | SARS-CoV-2 Antibody Calibrator | 20 μg |
| FCM04-C02 | RBD 6-plex Beads | 200 μL |
| FCM04-C03 | Assay Buffer | 55 mL |
| FCM04-C04 | 5 × Wash Buffer | 10 mL |
| FCM04-C05 | Biotinylated Human ACE2 | 40 μg |
| FCM04-C06 | 5 × PBS Buffer | 10 mL |
| FCM04-C07 | APC Beads | 500 μL |
| FCM04-C08 | PE Beads | 500 μL |
| FCM04-C09 | 96-Well V-bottom Plate | 1 plate |
| FCM04-C10 | 96-Well Sealing Film | 2 pieces |
| FCM04-C11 | SA-PE | 200 μL |
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) neutralizing 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 Neutralizing Antibody 6-plex Panel is a multiplex bead-based competitive immunofluorescence assay, utilizing flow cytometry for data acquisition and classification. Six SARS-CoV-2 Spike protein (WT / Alpha / Beta / Gamma / Delta / Omicron ) were conjugated to 6 fluorochrome-encoded microspheres respectively. Anti-SARS-CoV-2 neutralizing antibody in blood specimens competes with ACE2, for binding to Spike protein encapsulated 6-plex beads. Streptavidin conjugated R-phycoerythrin (SA-PE) was employed to detect biotinylated ACE2 bound to the beads. The intensity of PE fluorescence was assessed by flow cytometry at wavelength of 575 nm approximately, in inversely proportion to the Anti-SARS-CoV-2 neutralizing antibody in blood specimens. The intensity of APC fluorescence was applied to classify bead populations, at wavelength of 670 nm approximately.
活性（Bioactivity）-FACS Please refer to Ds document for the assay protocol.
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
A 4-parameter logistic model fitting curve was plotted on SARS-CoV-2 Variants Neutralizing Antibody 6-plex Panel. Immobilized SARS-CoV-2 variant Spike proteins bind with biotinylated human ACE2, to be competed out by a serial dilution of potent neutralizing antibody calibrator, ranging from 65 ng/ml to 16640 ng/ml. EC50 values indicate the concentration at which 50% of calibrator were bound. (QC tested).
Protocol
