OriGene validates every lot of every antibody it offers using Western blot analysis. We always assure that our antibodies detect the corresponding Gene Over Expression Cell Lysate - OriGene's Western Blot Standard (WBS) at the correct molecular weights in the Western blot analyses. OriGene provides an aliquot of the denatured Gene WBS lysate so the customer can verify gene expression using Western blot analysis.
The Western blot data is shown below for the p53 WBS (LY119832) together with the pCMV-XL5 vector transfected lysate validated by OriGene antibody (TA300033).
Second Level of Antibody Quality Control:
OriGene takes advantage of its large collection of TrueClone derived WBS lysates (see below) to select the antibody with the lowest cross-reactivity within the same protein families. For example, the OECLs for Akt family members (Akt1, Akt2 and Akt3), and GSK3 family members (GSK3a and GSK3ß) were used in Western blot analyses to assure high specificity. As seen from these data, OriGene provides antibodies with both high affinity and specificity.
Validated Gene Over Expression Cell Lysates - Western Blot Standard (WBS):
OriGene uses its TrueClone full length cDNA clones to generate WBS lysates. Each WBS lysate is validated against an antibody by detection of the corresponding protein at the correct molecular weight. To prepare WBS lysates, HEK293T cells are transiently transfected with 3 ug of TrueClone cDNA (in this case SC119832) using OriGene's optimal transfection reagent TurboFectin 8.0 (9 ul). Cells are lysed 72 hours post-transfection in a RIPA buffer. Protein concentration is determined using a colorimetric assay.