Human Mesothelin (296-580), His Tag (MSN-H5223) is expressed from human 293 cells (HEK293). It contains AA Glu 296 - Gly 580 (Accession # AAH09272).
Predicted N-terminus: Glu 296
This protein carries a polyhistidine tag at the C-terminus.
The protein has a calculated MW of 33.0 kDa. The protein migrates as 34-45 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
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.
Human Mesothelin (296-580), 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 95%.
The purity of Human Mesothelin (296-580), His Tag(Cat. No. MSN-H5223) was more than 90% and the molecular weight of this protein is around 45-60 kDa verified by SEC-MALS.
Immobilized Human Mesothelin (296-580), His Tag (Cat. No. MSN-H5223) at 0.5 μg/mL (100 μL/well) can bind Anti-Human MSLN MAb with a linear range of 0.1-2 ng/mL (QC tested).
Anti-Human MSLN MAb (chimeric mouse-human IgG1) captured on CM5 chip via Anti-Human IgG Fc antibodies surface, can bind Human Mesothelin (296-580), His Tag (Cat. No. MSN-H5223) with an affinity constant of 0.518 nM as determined in a SPR assay (Biacore T200) (Routinely tested).
Loaded Anti-Human MSLN MAb (chimeric mouse-human IgG1) on Protein A Biosensor, can bind Human Mesothelin (296-580), His Tag (Cat. No. MSN-H5223) with an affinity constant of 0.948 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely 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. Human Mesothelin (296-580), His Tag (Cat. No. MSN-H5223, 1 µg/mL) and C. His Tag protein control, followed by FITC-conjugated anti-6xHis tag antibody. A. Non-transfected 293 cells and C. His Tag-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 Human Mesothelin (296-580), His Tag (Cat. No. MSN-H5223).
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.