Human CD48 HEK293 Overexpression Lysate

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Human CD48 HEK293 Overexpression Lysate: Product Information

Product Description
This Human CD48 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD48 protein (Cat: 10797-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human CD48 (NP_001769.2) (Met 1-Ser 220) with a C-terminal polyhistidine tag was expressed.
Molecule Mass
The secreted recombinant human CD48 comprises 205 amino acids with a predicted molecular mass of 23.8 kDa. As a result of glycosylation, rhCD48 migrates as an approximately 40 kDa band in SDS-PAGE under reducing conditions.

Human CD48 HEK293 Overexpression Lysate: Usage Guide

Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
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.
Sample Buffer
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human CD48 HEK293 Overexpression Lysate: Alternative Names

Human BCM1 Overexpression Lysate; Human BLAST Overexpression Lysate; Human Blast-1 Overexpression Lysate; Human BLAST1 Overexpression Lysate; Human hCD48 Overexpression Lysate; Human mCD48 Overexpression Lysate; Human MEM-102 Overexpression Lysate; Human SLAMF2 Overexpression Lysate

CD48 Background Information

Cluster of Differentiation 48 (CD48), also known as SLAMF2, BCM-1 and BLAST-1, is a GPI-linked protein belonging to the CD2 subfamily of immunoglobulin superfamily molecules. CD2 and 2B4 (CD244) are known ligands for CD48. CD48 protein is expressed on most lineage-committed hematopoietic cells but not on hematopoietic stem cells or multipotent hematopoietic progenitors. CD48 protein performs biological functions in a variety processes including adhesion, pathogen recognition, cellular activation, and cytokine regulation, and emerges as a critical effector molecule in immune responses.
Full Name
CD48 molecule
References
  • Messmer B, et al. (2006) CD48 stimulation by 2B4 (CD244)-expressing targets activates human NK cells. J Immunol. 176(8): 4646-50
  • Milstein O, et al. (2008) Nanoscale increases in CD2-CD48-mediated intermembrane spacing decrease adhesion and reorganize the immunological synapse. J Biol Chem. 283(49): 34414-22.
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