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Your Position: Home > Immune Checkpoint Proteins > CTLA-4 > CT4-H52H9

Human CTLA-4 / CD152 Protein, His Tag, active dimer (MALS verified)

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    CTLA4,CD152
  • Source
    Human CTLA-4, His Tag (CT4-H52H9) is expressed from human 293 cells (HEK293). It contains AA Ala 37 - Phe 162 (Accession # Q6GR94-1).
    Predicted N-terminus: Ala 37
    Request for sequence
  • Molecular Characterization

    This protein carries a polyhistidine tag at the C-terminus.

    The protein has a calculated MW of 18.8 kDa. The protein migrates as 23-30 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

  • Endotoxin
    Less than 1.0 EU per μg by the LAL method.
  • Purity

    >90% as determined by SDS-PAGE.

    >90% as determined by SEC-MALS.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.

    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
Human CTLA-4, His Tag (Cat. No. CT4-H52H9) SDS-PAGE gel

Human CTLA-4, 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 90%.

SEC-MALS
Human CTLA-4, His Tag (Cat. No. ) MALS images

The purity of Human CTLA-4, His Tag (Cat. No. CT4-H52H9) was more than 90% and the molecular weight of this protein is around 45-60 kDa verified by SEC-MALS.

Report
Bioactivity-ELISA
Human CTLA-4, His TagHuman CTLA-4, His Tag (Cat. No. CT4-H52H9) ELISA bioactivity

Immobilized Human B7-1, Fc Tag (Cat. No. B71-H5259) at 2 μg/mL (100 μL/well) can bind Human CTLA-4, His Tag (Cat. No. CT4-H52H9) with a linear range of 0.1-2 ng/mL (QC tested).

Protocol
Human CTLA-4, His TagHuman CTLA-4, His Tag (Cat. No. CT4-H52H9) ELISA bioactivity

Immobilized Human CTLA-4, His Tag (Cat. No.CT4-H52H9) at 2 μg/mL, add increasing concentrations of CTLA-4 x OX40 Bispecific Antibody in 50% Human serum and then add Biotinylated Human OX40, Avitag,His Tag (Cat. No. TN4-H82E4) at 1 μg/mL. Detection was performed using HRP-conjugated streptavidin with sensitivity of 4 ng/mL (Intact assay, Routinely tested).

Protocol
Human CTLA-4, His TagHuman CTLA-4, His Tag (Cat. No. CT4-H52H9) ELISA bioactivity

Immobilized Human CTLA-4, His Tag (Cat. No. CT4-H52H9) at 1 μg/mL (100 μL/well) can bind Ipilimumab with a linear range of 0.1-3 ng/mL (Routinely tested).

Protocol
Bioactivity-BLI
Human_FcRn_Heterodimer_Protein_Bli

Loaded Human B7-1, Fc Tag (Cat. No. B71-H5259) on Protein A Biosensor, can bind Human CTLA-4, His Tag (Cat. No. CT4-H52H9) with an affinity constant of 0.498 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
Human_FcRn_Heterodimer_Protein_Bli

Loaded Human B7-2, Fc Tag (Cat. No. CD6-H5257) on Protein A Biosensor, can bind Human CTLA-4, His Tag (Cat. No. CT4-H52H9) with an affinity constant of 1.03 nM as determined in BLI assay (ForteBio Octet Red96e) (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. T cell activation through the T cell receptor and CD28 leads to increased expression of CTLA-4, an inhibitory receptor for B7 molecules. Fusion proteins of CTLA4 and antibodies (CTLA4-Ig) have been used in clinical trials for rheumatoid arthritis.
  • Clinical and Translational Updates
      

  • Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.

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