|20418C||1 Kit (96 assays)||
NOTE: This protocol is for PathScan® kits that use an HRP directly conjugated to the detection antibody (Rapid Protocol), rather than a 2-step method where the detection antibody and a secondary-HRP are added sequentially.
NOTE: Prepare solutions with deionized/purified water or equivalent.
For adherent cells
For suspension cells
NOTE: Equilibrate all materials and prepared reagents to room temperature prior to running the assay.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
created July 2020
Protocol Id: 2144
The poly-Glycine-Serine (G4S) linker is a type of flexible, unstructured synthetic peptide linker sequence often leveraged to connect the variable heavy (VH) domain and variable light (VL) domain of single-chain variable fragments (scFvs) and chimeric antigen receptors (CARs) that utilize an extracellular domain scFv for target antigen recognition. The linker itself consists of a core pentapeptide sequence, Gly-Gly-Gly-Gly-Ser, that is repeated and commonly found as either a 15-mer (G4S)3 or 20-mer (G4S)4 within scFv-based CARs and scFv fragments. The linker sequence length plays a role in controlling scFv stability and the noncovalent association between the VH and VL domains (1,2).
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