Biotinylated Human Glypican 3, His,Avitag (GP3-H82E5) is expressed from human 293 cells (HEK293). It contains AA Gln 25 - His 559 (Accession # P51654-1).
Predicted N-terminus: Gln 25 & Ser 359
This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag. This protein contains a furin-like convertase cleavage site, 355-RQYR-358, and will be partially processed into N and C-terminal fragment with calculated MW of 38.1 kDa and 26.4 kDa respectively. The protein migrates as 40 kDa and 70-120 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.
Less than 1.0 EU per μg by the LAL method.
>85% as determined by SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
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Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human Glypican 3, His,Avitag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 85%.
Immobilized Human FGF basic, Tag Free (Cat. No. BFF-H4117) at 2 μg/mL (100 μL/well) can bind Biotinylated Human Glypican 3, His,Avitag (Cat. No. GP3-H82E5) with a linear range of 1-16 ng/mL (QC tested).
Immobilized Anti-Human GPC3 Mab at 2 μg/mL (100 μL/well) can bind Biotinylated Human Glypican 3, His,Avitag (Cat. No. GP3-H82E5) with a linear range of 0.4-6 ng/mL (Routinely tested).
CAR阳性表达率检测（Evaluation of CAR expression ）
FACS Analysis of Anti-GPC3 CAR Expression
293 cells were transfected with anti-GPC3-scFv and RFP tag. 2e5 of the cells were first stained with B. Biotinylated Human Glypican 3, His,Avitag (Cat. No. GP3-H82E5, 10 μg/mL) and C. biotinylated protein control, followed by FITC-conjugated streptavidin. A. Non-transfected 293 cells and C. biotinylated protein control were used as negative control. RFP was used to evaluate CAR (anti-GPC3-scFv) expression and FITC was used to evaluate the binding activity of Biotinylated Human Glypican 3, His,Avitag (Cat. No. GP3-H82E5).
Authors: Berman RM, et al
Journal: Cancer Biother Radiopharm 43696
Glypican-3 (GPC3) is also known as Intestinal protein OCI-5, GTR2-2, MXR7, which belongs to the glypican family. Glypican 3 / GPC-3 is highly expressed in lung, liver and kidney. Glypican-3 inhibits the dipeptidyl peptidase activity of DPP4. Glypican-3 may be involved in the suppression/modulation of growth in the predominantly mesodermal tissues and organs, and also may play a role in the modulation of IGF2 interactions with its receptor and thereby modulate its function.