膜杰作
Star Staining
抗体来源（Source）
Monoclonal Anti-FMC63 Antibody, Rabbit IgG (1G10) is a rabbit monoclonal antibody recombinantly expressed from human 293 cells (HEK293), which provides higher batch consistency and long term security of supply. It shows superior performance in flow cytometry assays with higher affinity and better specificity.
应用（Application）
Flow Cytometry (Evaluation of Anti-CD19 (FMC63 scFv) CAR Expression).
克隆号（Clone）
1G10
种属（Species）
Rabbit
亚型（Isotype）
Rabbit IgG | Rabbit Kappa
特异性（Specificity）
Specifically recognizes the antigen-recognition domain of FMC63 derived CARs.
免疫原（Immunogen）
Recombinant FMC63 scFv derived from HEK293 cells.
偶联（Conjugate）
PE
Excitation Wavelength: 488 nm / 561 nm
Emission Wavelength: 575 nm
推荐稀释比（Recommended Dilution）
1:50
制剂（Formulation）
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4, 0.2% BSA, 0.03% Proclin300 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 24 months in lyophilized state;
- -70°C for 12 months after reconstitution.
- 2-8 °C for 12 month after reconstitution.
活性（Bioactivity）-FACS
Flow cytometric analysis of Anti-CD19 CAR-293 cells staining with PE-Labeled Monoclonal Anti-FMC63 Antibody, Rabbit IgG1 (1G10) (Cat. No. FM3-PFM721) at 1:50 dilution (2 μL of the antibody stock solution corresponds to labeling of 1e6 cells in a final volume of 100 µL), compared with isotype control antibody. PE signal was used to evaluate the binding activity (QC tested).
Protocol
Non-specificity of PE-Labeled Monoclonal Anti-FMC63 Antibody, Rabbit IgG1 (1G10) (Cat. No. FM3-PFM721) binding to CD3+ cells present in human PBMC. Human PBMCs were simultaneously stained with FITC-labeled anti-CD3 antibody and PE-Labeled Monoclonal Anti-FMC63 Antibody, Rabbit IgG1 (1G10) (2 μL of the antibody stock solution corresponds to labeling of 5e5 cells in a final volume of 100 µL), washed and then analyzed with FACS. Both FITC and PE positive signals was used to evaluate the non-specific binding activity to human CD3+ cells (Routinely tested).
Protocol
背景（Background）
FMC63 is an IgG2a mouse monoclonal antibody specific for CD19, which is a target for the immunotherapy of B lineage leukaemias and lymphomas. FMC63 scFv is the most commonly used ectodomain component of CD19-specific CARs. So far, most of reported CART19 trials contain the anti-CD19 scFv derived from FMC63, including the two FDA-approved CARs Kymriah and Yescarta.
