R Recombinant
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G4S Linker (E7O2V) Rabbit mAb (PE Conjugate) #38907
Filter:
- F
Flow cytometric analysis of a mixed population containing live wild-type Jurkat cells and Jurkat cells engineered to express an scFv-based Anti-CD20 CAR containing a G4S linker, using G4S Linker (E7O2V) Rabbit mAb (PE Conjugate) (right) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (PE Conjugate) #5742 (left). Tag Blue fluorescent protein (TagBFP) is co-expressed with the CAR. Cell line was provided by the Lohmueller Lab, University of Pittsburgh.
Supporting Data
| REACTIVITY | All |
| SENSITIVITY | Transfected Only |
| MW (kDa) | |
| Source/Isotype | Rabbit IgG |
Application Key:
- F-Flow Cytometry
Species Cross-Reactivity Key:
- All-All Species Expected
- Related Products
Product Information
Product Description
This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) fluorescent dye under optimal conditions and tested in-house for direct flow cytometric analysis in human cells. This antibody conjugate is expected to exhibit the same reactivity as the unconjugated G4S Linker (E7O2V) Rabbit mAb #71645, which is expected to react with cell surface-expressed CARs of varying specificity containing a G4S linker within the scFv of the extracellular domain.
Product Usage Information
| Application | Dilution |
|---|---|
| Flow Cytometry (Live) | 1:50 |
Storage
Supplied in PBS (pH 7.2), less than 0.1% sodium azide, and 2 mg/mL BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
Protocol
Specificity / Sensitivity
G4S Linker (E7O2V) Rabbit mAb (PE Conjugate) recognizes exogenously expressed levels of scFv-based CARs containing a G4S linker.
Species Reactivity:
All Species Expected
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide containing three Gly4Ser repeats.
Background
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).
限制使用
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For Research Use Only. Not For Use In Diagnostic Procedures.
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