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Mouse CD3D & CD3E Heterodimer HEK293 Cell Lysate (WB positive control)

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Mouse CD3D & CD3E Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Human Cells
Product Description:Human Cell lysate that Mouse CD3D & CD3E Heterodimer transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
Sequence information:A DNA sequence encoding the extracellular domain (Met 1-Ala 105) of mouse CD3D (P04235) was fused with a flag tag at the C-terminus, constructed the plasmid 1; A DNA sequence encoding the extracellular domain (Met 1-Asp 108) of mouse CD3E (P22646) was fused with a polyhistidine tag at the C-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the mouse CD3D/CD3E heterodimer was purified.
Predicted N Terminal:Phe 22 & Asp 22
Molecule Mass:The recombinant heterodimer of mouse CD3D/CD3E comprises 254 (126+128) amino acids and has a calculated molecular mass of 29.2 (14.3+14.9) KDa. As a result of glycosylation, the apparent molecular mass of mouse CD3D/CD3E heterodimer is approximately 20 and 22 KDa in SDS-PAGE under reducing conditions.
Mouse CD3D & CD3E Transfected / Overexpression Cell Lysate Usage Guide
Preparation Method:Cell lysate was prepared by homogenization 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.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
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. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
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Catalog: CT033-M2508HL-300
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