FITC-Labeled Human Glypican 3, His Tag (Cat. No. GP3-HF2H1) is expressed from human HEK293 cells. It contains AA Gln 25 - His 559 (Accession # P51654-1). It is the FITC labeled form of Human Glypican 3 (Cat. No. GP3-H52H4).
Predicted N-terminus: Gln 25
This protein carries a polyhistidine tag at the C-terminus.
The protein has a calculated MW of 62.7 kDa. The protein migrates as 32 kDa, 40 kDa and 66-115 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Excitation source: 488 nm spectral line, argon-ion laser
Excitation Wavelength: 488 nm
Emission Wavelength: 535 nm
The primary amines in the side chains of lysine residues and the N-terminus of the protein are conjugated with FITC using standard chemical labeling method. The residual FITC is removed by molecular seive treatment during purification process.
The FITC to protein molar ratio is 2.5-5.
Less than 1.0 EU per μg by the LAL method.
>90% 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 protect from light and avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
FITC-Labeled Human Glypican 3, His Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.
Immobilized Anti-hGPC3 antibody (Human IgG1) at 10 μg/mL (100 μL/well) can bind FITC-Labeled Human Glypican 3, His Tag (Cat. No. GP3-HF2H1) with a linear range of 0.078-0.625 μg/mL (QC 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 stained with B. FITC-Labeled Human Glypican 3, His Tag (Cat. No. GP3-HF2H1, 3 µg/ml) and C. FITC-labeled Protein Control. A. Non-transfected 293 cells and C. FITC-labeled 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 FITC-Labeled Human Glypican 3, His Tag (Cat. No. GP3-HF2H1).
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.