Human B7-1 HEK293 Overexpression Lysate: Product Information
This Human B7-1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of B7-1 protein (Cat: 10698-H03H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Asn 242) of human B7-1 (NP_005182.1) precursor was fused with the C-terminal His-tagged Fc region of human IgG1 at the C-terminus.
The recombinant human B7-1/Fc is a disulfide-linked homodimeric protein. The reduced monomer consists of 455 amino acids and predicts a molecular mass of 51.7 kDa. By SDS-PAGE, the apparent molecular mass of rh B7-1/Fc is approximately 80-90 kDa due to the glycosylation.
Human B7-1 HEK293 Overexpression Lysate: Usage Guide
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human B7-1 HEK293 Overexpression Lysate: Alternative Names
Human B7 Overexpression Lysate; Human B7-1 Overexpression Lysate; Human B7.1 Overexpression Lysate; Human BB1 Overexpression Lysate; Human CD28LG Overexpression Lysate; Human CD28LG1 Overexpression Lysate; Human LAB7 Overexpression Lysate
B7-1 Background Information
The B-lymphocyte activation antigen B7-1 (referred to as B7), also known as CD8, is a member of cell surface immunoglobulin superfamily and is expressed on the surface of antigen-presenting cells including activated B cells, macrophages and dendritic cells. As costimulatory ligands, B7-1 which exists predominantly as dimer and the related protein B7-2, interact with the costimulatory receptors CD28 and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) expressed on T cells, and thus constitute one of the dominant pathways that regulate T cell activation and tolerance, cytokine production, and the generation of CTL. The B7/CD28/CTLA4 pathway has the ability to both positively and negatively regulate immune responses. CD8 is thus regarded as promising therapeutic targets for autoimmune diseases and various carcinomas.
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