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Product Details
Source
Monoclonal Anti-FMC63 Antibody, Mouse IgG1, Avitag™ (Y45) is a neutralizing mouse monoclonal antibody recombinantly expressed from human 293 cells (HEK293). Biotinylation of this antibody is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
Antibody Type
Recombinant Monoclonal
Clone
Y45
Isotype
Mouse IgG1, Kappa
Host Species
Mouse
Immunogen
Recombinant FMC63 scFv derived from HEK293 cells
Conjugate
Biotin
Specificity
Specifically recognizes the antigen-recognition domain of FMC63 derived CARs.
Application
Flow Cytometry (Evaluation of Anti-CD19 (FMC63 scFv) CAR Expression).
Recommended Dilution
1:50
Isotype Control
The isotype control is sold separately; please refer to Cat. No. DNP-BM190 for product information.
Form
Lyophilized
Formulation
Lyophilized from a 0.22 μm-filtered solution in PBS (pH 7.4) containing 0.5% BSA, with trehalose as protectant.
Please contact us for customized product forms or formulations.
Reconstitution
Please refer to the Certificate of Analysis (CoA) for specific instructions.
Shipping
Lyophilized product is shipped at ambient temperature.
Storage
For long term storage, the product should be stored in a lyophilized state at -20°C or lower.
Please 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 under sterile conditions after reconstitution.
Notices
Product Specific Notices: For research use only.
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Performance Data
Bioactivity-FACS
2e5 of Anti-CD19 CAR-293 cells were stained with 100 μL of 1:50 dilution (2 μL stock solution in 100 μL FACS buffer) Biotinylated Monoclonal Anti-FMC63 Antibody, Mouse IgG1, Avitag (Cat. No. FM3-BY54) and isotype control respectively, washed and then followed by PE-SA and analyzed with FACS (QC tested).
Protocol
Non-specificity of Biotinylated Monoclonal Anti-FMC63 Antibody, Mouse IgG1, Avitag (Cat. No. FM3-BY54) binding to CD3+ cells present in human PBMC. Human PBMCs were simultaneously stained with FITC-labeled anti-CD3 antibody and Biotinylated Monoclonal Anti-FMC63 Antibody, Mouse IgG1, Avitag (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 (QC tested).
Protocol
FACS Analysis of Blood Samples
After Fc receptor blockade, whole blood samples (Figure A) and whole blood spiked with CAR-T cells (Figure B) were first stained with Biotinylated Monoclonal Anti-FMC63 Antibody, Mouse IgG1, Avitag™ (Cat. No. FM3-BY54). Following incubation with Streptavidin Protein-PE (Cat. No. STN-NP117) and 7-AAD staining, the cells were analyzed using FCS Express 7 software (Routinely tested). CAR-T cells were identified as PE-positive events within the live CD45+CD3+ lymphocyte population.
Protocol
Customer Reviews Writing Reviews
S***n2024-05-18
LikesWe have used this biotin anti-FMC63 for detecting our CD19 CAR on the surface of T cells. We use this in combination with a secondary staining with PE anti-Biotin at the recommended dilution and it has been working great. We have purchased multiple batches of this product and it has been consistent form lot to lot. We are happy with this and would recommend this for detecting FMC63.- S***i2024-05-10 LikesWe work with CD19 CAR T cells with the FMC63 scFv. We use this antibody to detect the surface expression of CD19 CAR on the T cells. We use a secondary PE anti-Biotin antibody following staining with this antibody. The surface staining using this antibody is bright and correlates with the secondary marker GFP we use. We have purchased multiple batches of this product and it has been reliable.
- F***e2022-07-15 LikesWe use this antibody for the selection of CAR cells from the immune cell population. It has been successful with the detection in in-vitro environments with FACS detection. The antibody can accurately select the positive population at a 1:200 dilution with very low background. The antibody is also very stable over long periods of time when it is stored correctly. I would recommend this product to anyone who is looking for a method for detection of FMC63.
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