Human CD28 HEK293 Overexpression Lysate: Product Information
This Human CD28 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD28 protein (Cat: 11524-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human CD28 isoform 1 (P10747-1) extracellular domain (Met 1-Pro 152) was was fused with the Fc region of human IgG1 at the C-terminus.
The recombinant human CD28/Fc is a disulfide-linked homodimeric protein. The reduced monomer consists of 375 amino acids and predictes a molecular mass of 42 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhCD28/Fc monomer is approximately 55-65 kDa due to glycosylation.
Human CD28 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 CD28 HEK293 Overexpression Lysate: Synonyms
Human Tp44 Overexpression Lysate
CD28 Background Information
CD28 (Cluster of Differentiation 28) is a disulphide-bonded glycoprotein belonging to the immunoglobulin (Ig) superfamily, and structurally consists of a single Ig V-like extracellular domain, a transmembrane domain and an intracellular domain. Mouse CD28 is constitutively expressed on the surface of all murine T cells and on developing thymocytes as disulfide-linked homodimers or as monomers. CD28 can binds the B7-1 and B7-2 ligand, and together perform important functions in the T and B cell response pathways. B7/CD28 family members, which can augment or antagonize T-cell receptor signaling, in the regulation of central and peripheral T-cell tolerance. CD28 is thus involved in T-cell activation, the induction of cell proliferation and cytokine production and promotion of T-cell survival.
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