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Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (Nanodisc) (HEK293)

分子别名(Synonym)

MS4A1,CD20,MS4A-1

表达区间及表达系统(Source)

Biotinylated Human CD20 Full Length, His,Avitag (CD0-H82E3) is expressed from human 293 cells (HEK293). It contains AA Met 1 - Pro 297 (Accession # P11836-1).

Predicted N-terminus: Met 1

蛋白结构(Molecular Characterization)

Online(Met 1 - Pro 297) P11836-1

Nanodiscs are a new class of model membranes that are being used to solubilize and study a range of integral membrane proteins (mp,full length CD20) and membrane-associated proteins. The Nanodisc bilayer is bounded by a membrane scaffold protein (MSP1D1) coat that confers enhanced stability and a narrow particle size distribution.
The CD20 nanodisc assembles from a mixture of full length CD20 protein in detergent, phospholipid micelles and membrane scaffold protein(MSP1D1) upon removal of the detergent.
The CD20 carries a polyhistidine tag at the C-terminus followed by an Avi tag with calculated MW of 36.9 kDa and migrates as 43 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation. The membrane scaffold protein(MSP1D1)has calculated MW of 24.7 kDa, and it migrates as 25 kDa under reducing (R) condition (SDS-PAGE) .

标记(Biotinylation)

Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.

内毒素(Endotoxin)

Less than 1.0 EU per μg by the LAL method.

纯度(Purity)

>90% as determined by SDS-PAGE.

制剂(Formulation)

Lyophilized from 0.22 μm filtered solution in 20 mM HEPES, 150 mM NaCl, pH7.5. Normally trehalose is added as protectant before lyophilization.

Contact us for customized product form or formulation.

重构方法(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.

存储(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. ● An empty/mock (no CD20) nanodisk is included in this product as negative control.
 

电泳(SDS-PAGE)

Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E3) SDS-PAGE gel

Biotinylated Human CD20 Full Length, 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 90%.

 

活性(Bioactivity)-ELISA

Biotinylated Human ELISA

Immobilized Rituximab at 5 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E3) with a linear range of 0.008-0.125 μg/mL (QC tested).

Protocol

活性(Bioactivity)-FACS

Biotinylated Human FACS

2e5 of CD20-CAR-293 cells transfected with anti-CD20-scFv were stained with 100 μL of 3 μg/mL of Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E3) and negative control protein respectively, washed and then followed by PE-SA and analyzed with FACS (QC tested).

Protocol

 

背景(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.

 

前沿进展

 
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CD20靶点信息
英文全称:B-lymphocyte antigen CD20
中文全称:B淋巴细胞抗原CD20
种类:Homo sapiens
上市药物数量:22详情
临床药物数量:72详情
最高研发阶段:批准上市
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