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G4S Linker (E7O2V) Rabbit mAb #71645
Filter:
- WB
- F
Simple Western™ analysis of lysates (1.5 mg/mL) from parental Jurkat cells, and Jurkat cells engineered to express an scFv-based Anti-CD19 CAR containing a G4S linker (Jurkat/CD19 CAR) using G4S Linker (E7O2V) Rabbit mAb #71645. The virtual lane view (left) shows the target band (as indicated) at a 1:50 dilution of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at a 1:50 dilution of primary antibody in parental Jurkat cells (black line) and Jurkat cells engineered to express an scFv-based Anti-CD19 CAR containing a G4S linker (Jurkat/CD19 CAR) (red line). This antibody is not approved for traditional Western blot. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module. Sample and stacking matrix load times were 6 seconds and 18 seconds, respectively. Baseline points were manually added to the electropherogram.
Supporting Data
| REACTIVITY | All |
| SENSITIVITY | Transfected Only |
| MW (kDa) | |
| Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- F-Flow Cytometry
Species Cross-Reactivity Key:
- All-All Species Expected
- Related Products
- Conjugates
Product Information
Product Description
This Cell Signaling Technology antibody is tested in-house for indirect flow cytometric analysis in human cells and is expected to react with cell surface expressed CARs of varying specificity, which contain a G4S linker within the scFv of the extracellular domain.
Product Usage Information
| Application | Dilution |
|---|---|
| Simple Western™ | 1:50 |
| Flow Cytometry (Live) | 1:50 - 1:200 |
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For a carrier free (BSA and azide free) version of this product see product #63670.
For a carrier free (BSA and azide free) version of this product see product #63670.
Protocol
Specificity / Sensitivity
G4S Linker (E7O2V) Rabbit mAb 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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