表达区间及表达系统(Source)
Biotinylated Human HLA-A*02:01&B2M&gp100 (YLEPGPVTA) Complex Protein (HLG-H82E6) is expressed from human 293 cells (HEK293). It contains AA Gly 25 - Thr 305 (HLA-A*02:01) & Ile 21 - Met 119 (B2M) & YLEPGPVTA peptide (Accession # AAA59606.1 (HLA-A*02:01) & P61769 (B2M) & YLEPGPVTA).
Predicted N-terminus: Gly 25 & Tyr
Request for sequence
蛋白结构(Molecular Characterization)
Biotinylated Human HLA-A*02:01&B2M&gp100 (YLEPGPVTA) Complex Protein is produced by co-expression of HLA and B2M loaded with gp100 peptide.
This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag (Avitag™).
The protein has a calculated MW of 36.0 kDa and 13.7 kDa. The protein migrates as 36-40 kDa, 17 kDa and 15 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
标记(Labeling)
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
蛋白标记度(Protein Ratio)
Passed as determined by the HABA assay / binding ELISA.
内毒素(Endotoxin)
Less than 1.0 EU per μg by the LAL method.
纯度(Purity)
>95% as determined by SDS-PAGE.
>95% as determined by SEC-MALS.
制剂(Formulation)
Lyophilized from 0.22 μm filtered solution in PBS, 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 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.
电泳(SDS-PAGE)
Biotinylated Human HLA-A*02:01&B2M&gp100 (YLEPGPVTA) Complex Protein on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.
SEC-MALS
The purity of Biotinylated Human HLA-A*02:01&B2M&gp100 (YLEPGPVTA) Complex Protein (Cat. No. HLG-H82E6) is more than 95% and the molecular weight of this protein is around 39-59 kDa verified by SEC-MALS.
Report
活性(Bioactivity)-ELISA
Immobilized Biotinylated Human HLA-A*02:01&B2M&gp100 (YLEPGPVTA) Complex Protein (Cat. No. HLG-H82E6) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-HLA class I Antibody, Human IgG1 (W6/32) with a linear range of 0.1-4 ng/mL (QC tested).
Protocol
Immobilized Biotinylated Human HLA-A*02:01&B2M&gp100 Protein (Cat. No. HLG-H82E6) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-B2M Antibody, Human IgG1 with a linear range of 0.1-2 ng/mL (Routinely tested).
Protocol
背景(Background)
Glycoprotein 100, is also well known as gp100 and Melanocyte protein PMEL. This antigen is a melanocyte-specific type I transmembrane glycoprotein and composed of 661 amino acids. Glycoprotein 100 is related to the tumor regression in metastatic melanoma patients after adoptive therapy, by which its epitopes being recognized by tumor lymphocytes and T-cells. The gp100280–288(YLEPGPVTA) was originally shown to be recognized by HLA-A*0201 tumor-infiltrating lymphocytes from melanoma patients, and therefore it is widely been studied in TCR -T studies. The Human HLA-A*0201 gp100 (YLEPGPVTA) complex protein is a complex of HLA-A*0201 of the MHC Class I, B2M and YLEPGPVTA peptide of the gp100.