表达区间及表达系统(Source)
Biotinylated Mouse H-2Kb&B2M&OVA (SIINFEKL) Complex Protein (H2A-M82E6) is expressed from human 293 cells (HEK293). It contains AA Gly 22 - Thr 305 (H-2Kb) & Ile 21 - Met 119 (B2M) & SIINFEKL peptide (Accession # P01901 (H-2Kb) & P01887 (B2M) & SIINFEKL).
Predicted N-terminus: Gly 22 & Ile 21
Request for sequence
蛋白结构(Molecular Characterization)
Biotinylated Mouse H-2Kb&B2M&OVA (SIINFEKL) Complex Protein is produced by co-expression of H-2Kb and B2M loaded with OVA peptide.
This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag (Avitag™).
The protein has a calculated MW of 39.7 kDa and 15.7 kDa. The protein migrates as 48-53 kDa and 17 kDa when calibrated against Star Ribbon Pre-stained Protein Marker 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.
>90% 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 Mouse H-2Kb&B2M&OVA (SIINFEKL) 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% (With Star Ribbon Pre-stained Protein Marker).
SEC-MALS
The purity of Biotinylated Mouse H-2Kb&B2M&OVA (SIINFEKL) Complex Protein (Cat. No. H2A-M82E6) is more than 90% and the molecular weight of this protein is around 55-70 kDa verified by SEC-MALS.
Report
活性(Bioactivity)-ELISA
Immobilized Biotinylated Mouse H-2Kb&B2M&OVA (SIINFEKL) Complex Protein (Cat. No. H2A-M82E6) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind anti-H-2Kb/OVA (SIINFEKL) antibody with a linear range of 0.1-4 ng/mL (QC tested).
Protocol
Immobilized Biotinylated Mouse H-2Kb&B2M&OVA (SIINFEKL) Complex Protein (Cat. No. H2A-M82E6) 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 0.1-1 ng/mL (Routinely tested).
Protocol
背景(Background)
Ovalbumin (OVA) can induce both humoral and cellular immune responses based on well-characterized peptide epitopes. The OVA257-264 octapeptide was one of the first OVA epitopes to be characterized, it has an amino acid sequence SIINFEKL, which is recognized by cytotoxic T lymphocytes. ovalbumin (Ova) was presented to T cells if it was released from pinosomes into the cytosol by osmotic lysis of pinosomes. Immunization with the adjuvanted SIINFEKL peptide induces long-lasting CD8+ T cell immunity in mice.