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- ProductSizeAmountActiveMax® Human CD19 μBeads, premium grade (for cells)2.5 mg2.5×10⁷ beadsActiveMax® Human CD19 μBeads, premium grade (for cells)10 mg (2.5 mg×4)1.0×10⁸ beads
产品描述（Product Description）
ActiveMax® Human CD19 μBeads, premium grade (for cells) are uniform, superparamagnetic beads of 5.5 µm in diameter immobilized with Human CD19 protein expressed from human 293 cells (HEK293) and contains AA Pro 20 - Lys 291 (Accession # P15391-1).
ActiveMax® Human CD19 μBeads, premium grade (for cells) are produced under sterile manufacturing conditions (ISO 5), and no animal- or human-derived components are used throughout the production process. It is produced under our rigorous quality control system that includes a comprehensive set of tests including sterility and endotoxin tests.应用说明（Application）
ActiveMax® Human CD19 μBeads, premium grade (for cells) are designed to stimulate in vitro CD19-specific CAR-T cells or UCAR-T cells, similar to the tumor cell lines that express human CD19 antigen. It can be used as follows:
Evaluating the characteristics of CAR-T cells or UCAR-T cells.
In vitro expansion of CD19-specific CAR-T cells or UCAR-T cells.
In vitro enrichment of CD19-specific CAR-T cells or UCAR-T cells.重构方法（Reconstitution）
See Certificate of Analysis (CoA) for detailed instruction.
存储（Storage）
This product is stable in storage under the following conditions: -20˚C for 12 months in lyophilized state. -70°C for 3 months under sterile conditions after reconstitution.
Please avoid repeated freeze-thaw cycles after reconstitution. Immediate use after reconstitution is highly recommended.
无菌（Sterility）
Negative
内毒素（Endotoxin）
Less than 0.002 EU per μg by the LAL method / rFC method.
注意事项（Important Note）
This product is for research use only and not intended for therapeutic or in vivo diagnostic use.
制剂（Formulation）
Please contact us for detailed information.
Contact us for customized product form or formulation.
质量管理控制体系（QMS）
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数据展示
流式数据 -FACS Data
Assay of human CD19 protein on the μBeads surface by Flow cytomtry. The human CD19 conjugated on the μBeads (Cat. No. MBS-C002) surface were fluorescently stained using PE labeled anti-human CD19 antibody and analyzed by flow cytometry (QC tested).
Protocol
批间一致性（Batch Consistency）
Add 100ul of 1:40 PE anti-human CD19 Antibody dilution (2.5ul stock solution in 100ul FACS buffer) into 5e5 of ActiveMax® human CD19 μBeads, premium grade (for cells) (Cat. No. MBS-C002), Negative control Beads (ActiveMax® Streptavidin μBeads, premium grade (for cells) (Cat. No. MBS-C009)) as well. PE signals was used to evaluate CD19 beads binding activity.
稳定性（Stability）
Add 100ul of 1:40 PE anti-human CD19 Antibody dilution (5ul stock solution in 100ul FACS buffer) into 5e5 of ActiveMax® human CD19 μBeads, premium grade (for cells) (Cat. No. MBS-C002), Negative control (ActiveMax® Streptavidin μBeads, premium grade (for cells) (Cat. No. MBS-C009)) as well. PE signals was used to evaluate CD19 beads binding activity.
刺激CD19 CAR-T细胞 -Cytokines
ActiveMax® Human CD19 μBeads, premium grade (for cells) (Cat. No. MBS-C002) can activate CD19-specific CAR-T cells by detecting the secretion of IFN- γ in vitro (Routinely tested).
CD19 CAR-T cells were stimulated with ActiveMax® Human CD19 μBeads (MBS-C002) for 24 h , and cell-free supernatants were harvested for evaluating IFN-γ secretion by ELISA. The results showed that CD19 CAR-T cells released significantly larger amounts of IFN-γ into the supernatants in response to CD19 μBeads .
Seed the CD19 μBeads in 96-well plates at a density of 2×10⁴ μBeads/well, and co-culture with CD19 CAR-T cells at an effector-to-target (E:T) ratio of 5:1, 1:1, 1:5 in 96-well tissue culture plate, respectively. CD19 CAR-T cells alone were cultured as the control.
CD19 CAR-T cells were stimulated with ActiveMax® Human CD19 μBeads (MBS-C002) for 24 h , and cell-free supernatants were harvested for evaluating IFN-γ secretion by flow cytomtry. The results showed that CD19 CAR-T cells released significantly larger amounts of IFN-γ into the supernatants in response to CD19 μBeads.
Seed the CD19 μBeads in 96-well plates at a density of 2×10⁴ μBeads/well, and co-culture with CD19 CAR-T cells at an effector-to-target (E:T) ratio of 5:1, 1:1, 1:5 in 96-well tissue culture plate, respectively. CD19 CAR-T cells alone were cultured as the control.
刺激CD19 CAR-T细胞 -Cytotoxic Molecules
CD19 CAR-T cells were stimulated with ActiveMax® Human CD19 μBeads (MBS-C002) for 24h, and cell-free supernatants were harvested for evaluating Granzyme B, Perforin secretion by ELISA. The results showed that CD19 CAR-T cells released significantly larger amounts of Granzyme B, Perforin into the supernatants in response to CD19 μBeads.
Seed the CD19 μBeads in 96-well plates at a density of 2×10⁴ μBeads/well, and co-culture with CD19 CAR-T cells at an effector-to-target (E:T) ratio of 5:1, 1:1, 1:5 in 96-well tissue culture plate, respectively. CD19 CAR-T cells alone were cultured as the control.
刺激CD19 CAR-T细胞 -Activation Marker
CD19 CAR-T cells were stimulated with ActiveMax® Human CD19 μBeads (MBS-C002) for 24h, and cells were harvested for evaluating CD69,CD137 expression by flow cytometry. The results showed that the proportion of CD69 and CD137 in CD19 CAR-T cells was significantly increased after CD19 μBeads stimulation.
Seed the CD19 μBeads in 96-well plates at a density of 2×10⁴ μBeads/well, and co-culture with CD19 CAR-T cells at an effector-to-target (E:T) ratio of 5:1, 1:1, 1:5 in 96-well tissue culture plate, respectively. CD19 CAR-T cells alone were cultured as the control.
刺激CD19 CAR-T细胞 -Exhaustion Marker
CD19 CAR-T cells were stimulated with ActiveMax® Human CD19 μBeads (MBS-C002) for 24h, and cells were harvested for evaluating Exhaustion Marker LAG-3, PD-1 expression by flow cytometry. The results showed that the proportion of LAG-3 and PD-1 in CD19 CAR-T cells was significantly increased after CD19 μBeads stimulation.
Seed the CD19 μBeads in 96-well plates at a density of 2×10⁴ μBeads/well, and co-culture with CD19 CAR-T cells at an effector-to-target (E:T) ratio of 5:1, 1:1, 1:5 in 96-well tissue culture plate, respectively. CD19 CAR-T cells alone were cultured as the control.
富集CD19 CAR-T细胞 -Low CAR Level
CD19 CAR-T cells with low gene expression were used as a starting point. The CAR positive cells were rare (~ 0.1%). After positive selection by the CD19 μBeads at a 2:4 (Beads:cells) ratio, the CAR positivity rate increased from an initial ~0.12% to 19%, corresponding to an approximately 150-fold enhancement.
Flow plots showing CD3+ CAR+ cells under magnetic enrichment with CD19 μBeads.
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