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Product Details
Source
Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade is a rabbit monoclonal antibody recombinantly expressed from human 293 cells (HEK293), it shows superior performance in various applications such as ELISA assays and flow cytometry assays with higher affinity and better specificity.
It is produced under our rigorous quality control system that incorporates a comprehensive set of tests including sterility and endotoxin tests. Product performance is carefully validated and tested for compatibility for cell culture use or any other applications in the early preclinical stage. When ready to transition into later clinical phases, we also offer a custom GMP protein service that tailors to your needs. We will work with you to customize and develop a GMP-grade product in accordance with your requests that also meets the requirements for raw and ancillary materials use in cell manufacturing of cell-based therapies.
Isotype
Rabbit IgG, Kappa
Conjugate
Biotin
Application
1. Flow Cytometry (Evaluation of cell surface expressed CARs of varying specificity containing a G4S linker within the scFv of the extracellular domain).
2. Immunoassay of bispecific antibody.
3. CAR T cell enrichment and sorting.
Specificity
It can specifically recognize proteins or ScFv containing (G3S)n or (G4S)n linkers, where n = 2, 3, 4... That is, it can recognize linkers with a minimum length of 2 repeats for both (G3S)n and (G4S)n.
Endotoxin
Less than 0.01 EU per μg by the LAL method / rFC method.
Sterility
Negative
Mycoplasma
Negative
Formulation
Lyophilized from 0.22 μm filtered solution in PBS, 0.2% rHSA, pH7.4 with trehalose as protectant.
Contact us for customized product form or formulation.
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.
Storage
For long term storage, the product should be stored at 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 3 months under sterile conditions after reconstitution.
ACRO Quality Management System
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Performance Data
Bioactivity-ELISA
Immobilized Recombinant Protein containing (G4S)2, Recombinant Protein containing (G4S)3 at 1 μg/mL (100 μL/well) can bind Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) with a linear range of 0.1-8 ng/mL (Routinely tested).
Protocol
Immobilized Recombinant Protein containing (G3S)2, Recombinant Protein containing (G3S)3, Recombinant Protein containing (G3S)4 at 1 μg/mL (100 μL/well) can bind Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) with a linear range of 0.1-31 ng/mL (Routinely tested).
Protocol
Bioactivity-FACS
Flow cytometric analysis of Anti-MSLN CAR-293 cells staining with Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) at 3 µg/mL (0.3 µg corresponds to labeling of 1e6 cells in a final volume of 100 µL) and PE-SA. PE signal was used to evaluate the binding activity (QC tested).
Protocol
Non-specificity of Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) binding to CD3+ cells present in human PBMC. 5e5 of human PBMCs were simultaneously stained with FITC-labeled anti-CD3 antibody and Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (0.3 µg corresponds to labeling of 5e5 cells in a final volume of 100 µL) and 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
Flow cytometric analysis of Anti-CD22 CAR-293 cells staining with Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) at 3 µg/mL (0.3 µg corresponds to labeling of 1e6 cells in a final volume of 100 µL) and PE-SA. PE signal was used to evaluate the binding activity (Routinely tested).
Protocol
Flow cytometric analysis of Anti-CD19 CAR-293 cells staining with Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) at 3 µg/mL (0.3 µg corresponds to labeling of 1e6 cells in a final volume of 100 µL) and PE-SA. PE signal was used to evaluate the binding activity (Routinely tested).
Protocol
Application Data
The Isolation of Anti-CD19 CAR-T cells with Biotinylated Anti-G4S Linker Antibody and SA Nanobeads.
Each 1E7 Anti-CD19 CAR-T cells were isolated with 0.6 µg Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) and appropriate SA Nanobeads. Both the isolated cells and non-isolated cells were stained with Alexa Fluor 647-Labeled Monoclonal Anti-FMC63 Antibody, Rabbit IgG (M4H08) (Cat. No. FM3-ABFMY2) and subsequently analyzed using FCS Express 7 software.
Following the separation process, the percentage of Anti-CD19 CAR-T cells was enriched from an initial 37.29% to a final purity of 96.37%, indicating a highly successful isolation.
The Isolation of Anti-BCMA CAR-T Cells with Biotinylated Anti-G4S Linker Antibody and SA Nanobeads.
Each 1E7 Anti-BCMA CAR-T cells were isolated with 0.6 µg Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) and appropriate SA Nanobeads. Both the isolated cells and non-isolated cells were stained with PE-labeled human BCMA/TNFRSF17 protein, His Tag, Star Staining (Cat. No. BCA-HP2H7) and subsequently analyzed using FCS Express 7 software.
Following the separation process, the percentage of Anti-BCMA CAR-T cells was enriched from an initial 32.99% to a final purity of 99.01%, indicating a highly successful isolation.
The Isolation of Anti-CD19 CAR 293 Cells with Biotinylated Anti-G4S Linker Antibody and SA Nanobeads.
Each 1E7 Anti-CD19 CAR 293 Cells were isolated with 0.2 µg Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) and appropriate SA Nanobeads. Both the isolated cells and non-isolated cells were stained with PE-Labeled Monoclonal Anti-FMC63 Antibody, Rabbit IgG (1G10) (0.03% Proclin) (Cat. No. FM3-PFM721) and subsequently analyzed using FCS Express 7 software.
Following the separation process, the percentage of Anti-CD19 CAR 293 cells was enriched from an initial 22.69% to a final purity of 96.01%, indicating a highly successful isolation.
The Isolation of Anti-CD19 CAR 293 Cells with Biotinylated Anti-G4S Linker Antibody and SA Nanobeads.
Each 1E7 Anti-CD19 CAR 293 Cells were isolated with 0.2 µg Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) and appropriate SA Nanobeads (Acro or Company M). The isolated cells were stained with PE-Labeled Monoclonal Anti-FMC63 Antibody, Rabbit IgG (1G10) (0.03% Proclin) (Cat. No. FM3-PFM721) and subsequently analyzed using FCS Express 7 software.
The results show that using Biotinylated Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016), premium grade (Cat. No. G4S-BLM25G0) combine with different SA nanobeads can all reach a final purity of over 95%, indicating a highly successful isolation.
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