Fresh or thawed T cells were activated for 24 hours on an OKT3 (1 μg/ml) and CD28 (1 μg/ml) antibody-coated 24-well plate in CTS OpTmizer medium (Gibco) supplemented with OpTmizer T cell expansion supplement, 10% FBS, 1% l -glutamine (200 mM), 1% penicillin (100 U/ml), 1% streptomycin (100 μg/ml), IL-7 (10 ng/ml), and IL-15 (5 ng/ml). Activated T cells were then transduced on non–tissue culture–treated plates coated with RetroNectin reagent (Takara Bio) (19 μg/ml) and CAR lentivirus (multiplicity of infection of approximately 10) according to the manufacturer’s instructions. CAR-T cells were then expanded for at least 7 days before use in experiments. Before all experiments, CAR-T cell transduction efficiency was normalized to 50% CAR+ cells (for in vitro cytotoxicity screening assay) or to the lowest efficiency donor by adding of non-transduced T cells. If necessary, CAR-T cells were purified using anti-phycoerythrin (PE) microbeads (Miltenyi Biotec) following anti-EGFR-PE staining according to the manufacturer’s instructions.
Anti phycoerythrin pe microbeads
Manufactured by Miltenyi Biotec
Sourced in Germany
Anti-phycoerythrin (PE) microbeads are a type of lab equipment used in flow cytometry and cell separation applications. These microbeads are coated with antibodies that specifically bind to the phycoerythrin fluorescent protein, allowing for the isolation or detection of cells or particles that express this protein.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using anti phycoerythrin pe microbeads
1
Lentiviral CAR-T Cell Generation
2
PEDV-induced Cytotoxicity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
On day 8 after piglets were challenged with PEDV, jejunal monocytes were stained using PE-labeled anti-pig CD8 (559584, BD, USA), and cells were subsequently collected as effector cells by positive screening in LS Columns (130-122-729, Miltenyi Biotec, Germany) using Anti-Phycoerythrin (PE) MicroBeads (130-048-801, Miltenyi Biotec, Germany). IPEC-J2 cells were grown to 70% confluence, the medium was discarded, and 10 μM LCA, 2 μM SBI-115 (HY-111534, MedChemExpress, USA), or 2 μM Gly-β-MCA (HY-114392, MedChemExpress, USA) was added. IPEC-J2 cells were subsequently infected with PEDV (103.5 PFU ml−1) for 8 h. After coculture of the effector and target cells for 4 h, the supernatant was collected, and the cytotoxicity of each group was assayed using the LDH Cytotoxicity Assay Kit (C0017, Beyotime, China) according to the manufacturer’s instructions.
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!