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Biotinylated Human Mesothelin / MSLN (296-580) Protein, His,Avitag™, premium grade DMF

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  • Synonym
    MSLN,Mesothelin,MPF
  • Source
    Biotinylated Human Mesothelin (296-580), His,Avitag, premium grade(MSN-H82E9) is expressed from human 293 cells (HEK293). It contains AA Glu 296 - Gly 580 (Accession # AAH09272.1).
    Predicted N-terminus: Glu 296
    It is produced under our rigorous quality control system that incorporates a comprehensive set of tests including sterility and endotoxin tests. Product performance is carefully validated and tested for compatibility for cell culture use or any other applications in the early preclinical stage. When ready to transition into later clinical phases, we also offer a custom GMP protein service that tailors to your needs. We will work with you to customize and develop a GMP-grade product in accordance with your requests that also meets the requirements for raw and ancillary materials use in cell manufacturing of cell-based therapies.
  • Molecular Characterization
    Mesothelin Structure

    This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag (Avitag™).

    The protein has a calculated MW of 35.9 kDa. The protein migrates as 40-50 kDa when calibrated against Star Ribbon Pre-stained Protein Marker under reducing (R) condition (SDS-PAGE) due to glycosylation.

  • Labeling
    Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
  • Protein Ratio
    Passed as determined by the HABA assay / binding ELISA.
  • Endotoxin
    Less than 0.01 EU per μg by the LAL method.
  • Sterility
    Negative
  • Mycoplasma
    Negative.
  • Purity

    >95% as determined by SDS-PAGE.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.

    Contact us for customized product form or formulation.

  • Reconstitution

    Please see Certificate of Analysis for specific instructions.

    For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

  • Storage

    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:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 3 months under sterile conditions after reconstitution.
SDS-PAGE
Mesothelin SDS-PAGE

Biotinylated Human Mesothelin (296-580), His,Avitag, premium grade on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95% (With Star Ribbon Pre-stained Protein Marker).

SEC-MALS
Mesothelin MALS images

The purity of Biotinylated Human Mesothelin (296-580), His,Avitag, premium grade (Cat. No. MSN-H82E9) is more than 85% and the molecular weight of this protein is around 38-53 kDa verified by SEC-MALS.

Bioactivity-FACS
 Mesothelin FACS

2e5 of Anti-MSLN CAR-293 cells were stained with 100μL of 1 μg/mL of Biotinylated Human Mesothelin (296-580), His,Avitag, premium grade (Cat. No. MSN-H82E9) and negative control protein respectively, washed and then followed by PE-SA and analyzed with FACS (QC tested).

Bioactivity-ELISA
 Mesothelin ELISA

Immobilized Anti-Human MSLN MAb at 0.5 μg/mL (100 μL/well) can bind Biotinylated Human Mesothelin (296-580), His,Avitag, premium grade (Cat. No. MSN-H82E9) with a linear range of 0.1-0.78 ng/mL (QC tested).

Evaluation of CAR expression
FACS Analysis of Anti-MSLN CAR Expression
 Mesothelin CAR_T

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, premium grade (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, premium grade (Cat. No. MSN-H82E9).

  • Background
    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.
  • Clinical and Translational Updates

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