Human CD8 alpha HEK293 Overexpression Lysate: Product Information
This Human CD8 alpha overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD8 alpha protein (Cat: 10980-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of human CD8A (NP_001759.3) (Met 1-Asp 182) was expressed with a C-terminal polyhistidine tag.
The recombinant human CD8A consists of 172 amino acids and predicts a molecular mass of 19 kDa. As a result of glycosylation, rh CD8A migrates as an approximately 28 kDa band in SDS-PAGE under reducing conditions.
Human CD8 alpha 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 CD8 alpha HEK293 Overexpression Lysate: Alternative Names
Human CD8 Overexpression Lysate; Human CD8 alpha Overexpression Lysate; Human Leu-2 Overexpression Lysate; Human Leu2 Overexpression Lysate; Human MAL Overexpression Lysate; Human p32 Overexpression Lysate; Human p32 186910 Overexpression Lysate
CD8 alpha Background Information
T-cell surface glycoprotein CD8 alpha chain, also known as CD8a, is a single-pass type I membrane protein. The CD8 glycoprotein is expressed by thymocytes, mature T cells and natural killer (NK) cells and has been implicated in the recognition of monomorphic determinants on major histocompatibility complex (MHC) Class I antigens, and in signal transduction during the course of T-cell activation. Both human and rodent CD8 antigens are comprised of two distinct polypeptide chains, alpha and beta. The Ig domains of CD8 alpha are involved in controlling the ability of CD8 to be expressed. Mutation of B- and F-strand cysteine residues in CD8 alpha reduced the ability of the protein to fold properly and, therefore, to be expressed. Defects in CD8A are a cause of familial CD8 deficiency. Familial CD8 deficiency is a novel autosomal recessive immunologic defect characterized by absence of CD8+ cells, leading to recurrent bacterial infections.
References Devine, L. et al., 2000, J Immunol. 164 (2): 833-8. Arcaro, A. et al., 2000, J Immunol. 165 (4): 2068-76. Saha, K. et al., 2001, Nat Med. 7 (1): 65-72. Romero, P. et al., 2005, Eur J Immunol. 35 (11): 3092-4.
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