Biotinylated Human Mesothelin (296-580), His,Avitag (MSN-H82E9) is expressed from human 293 cells (HEK293). It contains AA Glu 296 - Gly 580 (Accession # AAH09272.1).
Predicted N-terminus: Glu 296
This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag.
The protein has a calculated MW of 35.9 kDa. The protein migrates as 45-60 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.
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.
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 Mesothelin (296-580), 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 95%.
The purity of Biotinylated Human Mesothelin (296-580), His,Avitag(Cat. No. MSN-H82E9) was more than 90% and the molecular weight of this protein is around 45-55 kDa verified by SEC-MALS.
Immobilized Anti-Human MSLN MAb at 0.5 μg/mL (100 μL/well) can bind Biotinylated Human Mesothelin (296-580), His,Avitag (Cat. No. MSN-H82E9) with a linear range of 0.1-0.78 ng/mL (QC tested).
CAR阳性表达率检测（Evaluation of CAR expression ）
FACS Analysis of Anti-MSLN CAR Expression
293 cells were transfected with Anti-MSLN-scFv and RFP tag. 2e5 of the cells were first stained with B. Biotinylated Human Mesothelin (296-580), His,Avitag (Cat. No. MSN-H82E9, 3 µg/mL) and C. Biotinylated Protein Control, followed by FITC Streptavidin. A. Non-transfected 293 cells and C. Biotinylated Protein Control were used as negative control. RFP was used to evaluate CAR (Anti-MSLN-scFv) expression and FITC was used to evaluate the binding activity of Biotinylated Human Mesothelin (296-580), His,Avitag (Cat. No. MSN-H82E9).
Mesothelin (MSLN) is also known as CAK1 antigen, Pre-pro-megakaryocyte-potentiating factor, which belongs to the mesothelin family. Mesothelin / MSLN can be proteolytically cleaved into the following two chains by a furin-like convertase: Megakaryocyte-potentiating factor (MPF) and the cleaved form of mesothelin. Both MPF and the cleaved form of mesothelin are N-glycosylated. Mesothelin / MSLN can interacts with MUC16. The membrane-anchored forms of MSLN may play a role in cellular adhesion. MPF potentiates megakaryocyte colony formation in vitro.