For detection of antigen-specific B cells, spike protein (R&D Systems) was biotinylated using the biotin conjugation kit (Abcam), and was subsequently labeled for 1 hour at 4 °C with streptavidin (SA)-allophycocyanin (APC)/phycoerythrin (PE) (Miltenyi Biotec) at a 5:1 mass ratio and with SA-FITC (Miltenyi Biotec) at a 4:1 mass ratio, respectively. Ten million resting PBMCs and SA-fluorescence-labeled biotinylated proteins (protein probes) were treated with 5 mM of d-biotin (Cayman Chemical) for 15 min at room temperature, and then stained with protein probes (0.6 μg spike-PE/APC; 0.4 μg receptor binding domain derived from spike protein (RBD-FITC) for 30 min. The RBD-FITC was from Dr. Mi-Hua Tao at Academia Sinica, Taipei, Taiwan. Cells were then washed twice with staining buffer, and stained by a panel of surface marker antibodies, including anti-CD3 PE-Cy7, anti-CD19 BV421, anti-CD20 PE-CF594, anti-IgG APC-H7, and anti-IgM BUV395. Finally, cells were analyzed using BD FACS LSRFortessa™ cytometer.
Spike protein
Manufactured by R&D Systems
The Spike protein is a laboratory product used in research and development. It is a structural component of the SARS-CoV-2 virus, which causes COVID-19. The Spike protein plays a critical role in the virus's ability to bind to and infect human cells. This product can be used in various research applications, such as studying virus-host interactions, vaccine development, and diagnostic test development.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using spike protein
1
Detecting Antigen-Specific B Cells
2
SARS-CoV-2 Spike Protein Antibody Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The levels of IgG and IgA antibodies against Spike Protein were determined by ELISA by sensitizing the plate with homogenates of killed whole virus produced in vitro. Spike was quantified using purified protein (Human coronavirus HCoV-229E Spike Protein (S1+S2 ECD). Sino Biological, Inc. 40605-V08B and Spike Protein (Active Trimer) R&D Cat. # 10549-CV). The following secondary antibodies were used: Mouse anti-Hamster IgG Cocktail, Clone: G94-56, G70-204 (BD Biosciences, Cat. # 554009); Mouse anti-Hamster IgM, Clone: G188-9, (BD Biosciences Cat. # 554035); Hamster Immunoglobulin A (IgA) ELISA Kit (MyBiosources Cat. # MBS029668); Mouse monoclonal (H6) anti-SARS-CoV-2 spike glycoprotein (Abcam Cat. # ab273169).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!