膜杰作
Star Staining
分子别名（Synonym）
HLA-A*0201 & B2M
表达区间及表达系统（Source）
PE-Labeled Human HLA-A*02:01&B2M Tetramer Protein (HLM-HP2H3) is expressed from human 293 cells (HEK293). It contains AA Gly 25 - Ile 308 (HLA-A*02:01) & Ile 21 - Met 119 (B2M) (Accession # AAA59606.1 (HLA-A*02:01) & P61769-1 (B2M)).
Predicted N-terminus: Gly 25 & Ile 21
Request for sequence
蛋白结构（Molecular Characterization）
PE-Labeled Human HLA-A*02:01&B2M Tetramer Protein is assembled by biotinylated monomer (HLM-H82W3) and PE-labeled streptavidin.
Biotinylated Human HLA-A*02:01&B2M Complex Protein is produced by co-expression of HLA and B2M. This Protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag (Avitag™).
偶联（Conjugate）
PE
Excitation Wavelength: 488 nm / 561 nm
Emission Wavelength: 575 nm
内毒素（Endotoxin）
Less than 1.0 EU per μg by the LAL method.
纯度（Purity）
>90% as determined by SDS-PAGE.
制剂（Formulation）
Lyophilized from 0.22 μm filtered solution in PBS, 1% 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 3 months under sterile conditions after reconstitution.
活性（Bioactivity）-ELISA
Immobilized PE-Labeled Human HLA-A*02:01&B2M Tetramer Protein (Cat. No. HLM-HP2H3) at 1 μg/mL (100 μL/well) can bind Anti-HLA class I Antibody, Human IgG1 (W6/32) with a linear range of 0.1-2 ng/mL (QC tested).
Protocol
Immobilized PE-Labeled Human HLA-A*02:01&B2M Tetramer Protein (Cat. No. HLM-HP2H3) at 1 μg/mL (100 μL/well) can bind Anti-B2M Antibody, Human IgG1 with a linear range of 0.1-1 ng/mL (Routinely tested).
Protocol
CAR阳性表达率检测（Evaluation of CAR expression）
FACS Analysis of Non-specific binding to PBMCs
Non-specificity of PE-Labeled Human HLA-A*02:01&B2M Tetramer Protein (Cat. No. HLM-HP2H3) binding to CD4+ cells present in human PBMC. 5e5 of human PBMCs were simultaneously stained with Alexa Fluor® 700 anti-human CD4 Antibody and PE-Labeled Human HLA-A*02:01&B2M Tetramer Protein (1 µg corresponds to labeling of 5e5 cells in a final volume of 100 µL) and washed and then analyzed with FACS. Both Alexa 700 and PE positive signals was used to evaluate the non-specific binding activity to human CD4+ cells (Routinely tested).
FACS Analysis of Non-specific binding to PBMCs
Non-specificity of PE-Labeled Human HLA-A*02:01&B2M Tetramer Protein (Cat. No. HLM-HP2H3) binding to CD8+ cells present in human PBMC. 5e5 of human PBMCs were simultaneously stained with FITC anti-human CD8 Antibody and PE-Labeled Human HLA-A*02:01&B2M Tetramer Protein (1 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 CD8+ cells (Routinely tested).
