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Your Position: ホーム > Protein > CD20 > CD0-H82E5

Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (Detergent) (SPR verified)

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    MS4A1,CD20,MS4A-1
  • Source
    Biotinylated Human CD20 Full Length, His,Avitag(CD0-H82E5) is expressed from human 293 cells (HEK293). It contains AA Met 1 - Pro 297 (Accession # P11836-1).
    Predicted N-terminus: Met 1
    Request for sequence
  • Molecular Characterization
    CD20 Structure

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

    The protein has a calculated MW of 36.9 kDa. The protein migrates as 44 kDa 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 1.0 EU per μg by the LAL method.
  • Purity

    >90% as determined by SDS-PAGE.

  • Formulation

    This product is not suitable for cell based experiments due to cytotoxicity of DDM.
    DDM and CHS are INDISPENSABLE to keep membrane protein soluble and active, under no circumastance should you remove DDM and CHS.
    DDM/CHS buffer (DC-11) is sold separately and not included in protein, and please contact us if you need the buffer.
    If glycerol is not compatible to your application, remove glycerol just before immediate experiment, and NEVER store glycerol-free protein solution.

    Supplied as 0.2 μm filtered solution in 50 mM HEPES, 150 mM NaCl, DDM, CHS, pH7.5 with glycerol as protectant.

    Contact us for customized product form or formulation.

  • Shipping

    This product is supplied and shipped with dry ice, please inquire the shipping cost.

  • Storage

    Please avoid repeated freeze-thaw cycles.

    This product is stable after storage at:

    1. The product MUST be stored at -70°C or lower upon receipt;
    2. -70°C for 3 months under sterile conditions.
*The DDM/CHS buffer (Cat. No. DC-11) is sold separately and not included in protein, you can follow this link for product information.
SDS-PAGE
CD20 SDS-PAGE

Biotinylated Human CD20 Full Length, His,Avitag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90%.

Bioactivity-ELISA
CD20 ELISA

Immobilized Rituximab biosimilar (Cat. No. CD0-M36) at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) with a linear range of 1-15.625 ng/mL (QC tested).

Protocol
CD20 ELISA

Immobilized Ofatumumab at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) with a linear range of 4-63 ng/mL (in presence of DDM and CHS) (QC tested).

Protocol
CD20 ELISA

Immobilized Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Obinutuzumab with a linear range of 1.2-2 ng/mL (Routinely tested).

Protocol
CD20 ELISA

Immobilized Human CD3E&CD3D Heterodimer Protein (Cat. No. CDD-H52Wa) at 1 μg/mL, add Glofitamab and then add Biotinylated Human CD20 Protein (Cat. No. CD0-H82E5) at 0.1 μg/mL. Detection was performed using HRP-conjugated Streptavidin (Acro, Cat. No. STN-NH913) (Routinely tested).

Protocol
Bioactivity-SPR
CD20 SPR

Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) captured on Biotin CAP-Series S Sensor Chip can bind Rituximab with an affinity constant of 1.73 nM as determined in a SPR assay (in presence of DDM and CHS) (Biacore T200) (QC tested).

Protocol
CD20 SPR

Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) captured on Biotin CAP-Series S Sensor Chip can bind Ofatumumab (Human IgG1) with an affinity constant of 1.03 nM as determined in a SPR assay (in presence of DDM and CHS) (Biacore T200) (QC tested).

Protocol
CD20 SPR

Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) captured on Biotin CAP-Series S Sensor Chip can bind Obinutuzumab with an affinity constant of 0.721 nM as determined in a SPR assay (in presence of DDM and CHS) (Biacore T200) (Routinely tested).

Protocol
Bioactivity-BLI
CD20 BLI

Loaded Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) on SA Biosensor, can bind Rituximab with an affinity constant of 0.247 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
CD20 BLI

Loaded Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) on SA Biosensor, can bind Ofatumumab (Human IgG1) with an affinity constant of 0.277 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
CD20 BLI

Loaded Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) on SA Biosensor, can bind Biosimilar of Obinutuzumab with an affinity constant of 0.197 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
  • Background
    B-lymphocyte antigen CD20 is also known as B-lymphocyte surface antigen B1, Leukocyte surface antigen Leu-16, Membrane-spanning 4-domains subfamily A member 1 and MS4A1, is an activated-glycosylated phosphoprotein expressed on the surface of all B-cells beginning at the pro-B phase (CD45R+, CD117+) and progressively increasing in concentration until maturity. CD20 is expressed on all stages of B cell development except the first and last; it is present from late pro-B cells through memory cells, but not on either early pro-B cells or plasma blasts and plasma cells. It is found on B-cell lymphomas, hairy cell leukemia, B-cell chronic lymphocytic leukemia, and melanoma cancer stem cells. The protein has no known natural ligand and its function is to enable optimal B-cell immune response, specifically against T-independent antigens. It is suspected that it acts as a calcium channel in the cell membrane. CD20 / MS4A1 is the target of the monoclonal antibodies (mAb) rituximab, Ibritumomab tiuxetan, and tositumomab, which are all active agents in the treatment of all B cell lymphomas and leukemias. Defects in CD20 / MS4A1 are the cause of immunodeficiency common variable type 5 (CVID5); also called antibody deficiency due to CD20 defect. CVID5 is a primary immunodeficiency characterized by antibody deficiency, hypogammaglobulinemia, recurrent bacterial infections and an inability to mount an antibody response to antigen.
  • Clinical and Translational Updates

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