抗体来源(Source)
Monoclonal Anti-G4S linker Antibody, Mouse IgG1 (AM635) is a mouse monoclonal antibody recombinantly expressed from human 293 cells (HEK293), which provides higher batch consistency and long term security of supply. APC-Labeled Monoclonal Anti-G4S linker Antibody, Mouse IgG1 (AM635) is produced via site-specific conjugation of APC to Monoclonal Anti-FMC63 Antibody, Mouse IgG1 under optimal conditions with a proprietary technology.
应用(Application)
Flow Cytometry (Evaluation of cell surface expressed CARs of varying specificity containing a G4S linker within the scFv of the extracellular domain). Please note that this product is NOT compatible to streptavidin detection system.
克隆号(Clone)
AM635
种属(Species)
Mouse
亚型(Isotype)
Mouse IgG1/kappa
特异性(Specificity)
Specifically recognizes the scFv-based CARs containing a G4S linker.
偶联(Conjugate)
APC
Excitation Wavelength: 640 nm
Emission Wavelength: 661 nm
推荐稀释比(Recommended Dilution)
1:25
制剂(Formulation)
Lyophilized from 0.22 μm filtered solution in PBS, 0.5% BSA, pH7.4 with trehalose as protectant.
Contact us for customized product form or formulation.
重构方法(Reconstitution)
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
存储(Storage)
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please protect from light and avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 12 months under sterile conditions after reconstitution.
活性(Bioactivity)-FACS
![G4S linker FACS](images/facs/G4S-AFM664-F1.jpg)
Flow cytometric analysis of Anti-MSLN CAR-293 cells staining with APC-Labeled Monoclonal Anti-G4S linker Antibody, Mouse IgG1(AM635) (Cat. No. G4S-AFM664) at 1:25 dilution(4 μL of the antibody stock solution corresponds to labeling of 1e6 cells in a final volume of 100 µL) , compared with isotype control antibody. APC signal was used to evaluate the binding activity (QC tested).
Protocol
![G4S linker FACS](images/facs/G4S-AFM664-F2.jpg)
Non-specificity of APC-Labeled Monoclonal Anti-G4S linker Antibody, Mouse IgG1(AM635) (Cat. No. G4S-AFM664) binding to CD3+ cells present in human PBMC. 5e5 of human PBMCs were simultaneously stained with FITC-labeled anti-CD3 antibody and APC-Labeled Monoclonal Anti-G4S linker Antibody (4 μL of the antibody stock solution corresponds to labeling of 5e5 cells in a final volume of 100 µL) and washed and then analyzed with FACS. Both FITC and APC positive signals was used to evaluate the non-specific binding activity to human CD3+ cells (QC tested).
Protocol
![G4S linker FACS](images/facs/G4S-AFM664-F3.jpg)
Flow cytometric analysis of Anti-CD19 CAR-293 cells staining with APC-Labeled Monoclonal Anti-G4S linker Antibody, Mouse IgG1(AM635) (Cat. No. G4S-AFM664) at 1:25 dilution(4 μL of the antibody stock solution corresponds to labeling of 1e6 cells in a final volume of 100 µL) , compared with isotype control antibody. APC signal was used to evaluate the binding activity(Routinely tested).
Protocol
![G4S linker FACS](images/facs/G4S-AFM664-F4.jpg)
Flow cytometric analysis of HEK293 cells staining with APC-Labeled Monoclonal Anti-G4S linker Antibody, Mouse IgG1(AM635) (Cat. No. G4S-AFM664) at 1:25 dilution(4 μL of the antibody stock solution corresponds to labeling of 1e6 cells in a final volume of 100 µL) , compared with isotype control antibody. APC signal was used to evaluate the binding activity(Routinely tested).
Protocol
背景(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 or20-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.