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Your Position: ホーム > Protein > CTLA-4 > CT4-H82E3

Biotinylated Human CTLA-4 Protein, His,Avitag™ (MALS verified)

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    CTLA4,CD152
  • Source
    Biotinylated Human CTLA-4, His,Avitag(CT4-H82E3) is expressed from human 293 cells (HEK293). It contains AA Ala 37 - Phe 162 (Accession # P16410-1).
    Predicted N-terminus: Ala 37
    Request for sequence
  • Molecular Characterization
    CTLA-4 Structure

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

    The protein has a calculated MW of 17.2 kDa. The protein migrates as 22-30 kDa under reducing (R) condition, and 37-45 kDa under non-reducing (NR) 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

    >95% as determined by SDS-PAGE.

    >90% as determined by SEC-MALS.

  • 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
CTLA-4 SDS-PAGE

Biotinylated Human CTLA-4, His,Avitag on SDS-PAGE under reducing (R) and non-reducing (NR) conditions. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.

SEC-MALS
CTLA-4 MALS images

The purity of Biotinylated Human CTLA-4, His,Avitag (Cat. No. CT4-H82E3) is more than 90% and the molecular weight of this protein is around 40-60 kDa verified by SEC-MALS.

Report
Bioactivity-ELISA
CTLA-4 ELISA

Immobilized Human B7-1, Fc Tag (Cat. No. B71-H5259) at 2 μg/mL (100 μL/well) can bind Biotinylated Human CTLA-4, His,Avitag (Cat. No. CT4-H82E3) with a linear range of 0.7-13 ng/mL (QC tested).

Protocol
CTLA-4 ELISA

Quantitative Analysis of CTLA-4 x OX40 Bispecific Antibody in Human Serum by Intact Assay.

Immobilized Human OX40 Protein, His Tag (MALS verified) (Cat. No. OX0-H5224) at 2 μg/mL, add increasing concentrations of CTLA-4 x OX40 bispecific antibody in 50% Human serum and then add Biotinylated Human CTLA-4, His, Avitag (Cat. No. CT4-H82E3) at 0.2 μg/mL. Detection was performed using HRP-conjugated streptavidin with sensitivity of 4 ng/mL (Intact assay, Routinely tested).

Protocol
Bioactivity-SPR
CTLA-4 SPR

Biotinylated Human CTLA-4, His,Avitag (Cat. No. CT4-H82E3) captured on Biotin CAP - Series S sensor Chip can bind Yervoy (Ipilimumab) with an affinity constant of 0.635 nM as determined in a SPR assay (Biacore T200) (Routinely tested).

Protocol
  • Background
    CTLA-4 (Cytotoxic T-Lymphocyte Antigen 4) is also known as CD152 (Cluster of differentiation 152), is a protein receptor that downregulates the immune system. CTLA4 is a member of the immunoglobulin superfamily, which is expressed on the surface of Helper T cells and transmits an inhibitory signal to T cells. The protein contains an extracellular V domain, a transmembrane domain, and a cytoplasmic tail. Alternate splice variants, encoding different isoforms. CTLA4 is similar to the T-cell co-stimulatory protein, CD28, and both molecules bind to CD80 and CD86, also called B7-1 and B7-2 respectively, on antigen-presenting cells. CTLA4 transmits an inhibitory signal to T cells, whereas CD28 transmits a stimulatory signal. Intracellular CTLA4 is also found in regulatory T cells and may be important to their function. Fusion proteins of CTLA4 and antibodies (CTLA4-Ig) have been used in clinical trials for rheumatoid arthritis.
  • Clinical and Translational Updates

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