Biotinylated Recombinant Human CTLA-4-Fc Chimera (carrier-free)

Pricing & Availability
Regulatory Status
RUO
Other Names
CD152, CELIAC3, GSE, IDDM12, cytotoxic T-lymphocyte-associated protein 4, ALPS5, and GRD4
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Human__CTLA-4_Chimera-CF_BIOTIN_RECOM_1_091719
When human CD86-Fc Chimera (Cat. No. 775602) is immobilized at 1.0 µg/mL, Biotinylated Recombinant Human CTLA-4-Fc Chimera binds with an EC50 of 12 - 48 ng/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding.
  • Human__CTLA-4_Chimera-CF_BIOTIN_RECOM_1_091719
    When human CD86-Fc Chimera (Cat. No. 775602) is immobilized at 1.0 µg/mL, Biotinylated Recombinant Human CTLA-4-Fc Chimera binds with an EC50 of 12 - 48 ng/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding.
  • Human__CTLA-4_Chimera-CF_BIOTIN_RECOM_2_091719
    Stability testing for Biotinylated Recombinant Human CTLA-4-Fc Chimera. Recombinant human CTLA-4-Fc Chimera Biotin was aliquoted in PBS, pH 7.2, 5% glycerol at 0.2 mg/mL. One aliquot was frozen and thawed four times (4x Freeze/Thaw), and compared to a control kept at 4°C (Control). The samples were tested in a binding assay with Recombinant Human B7-2/CD86-Fc Chimera (Cat. No. 775602).
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786702 10 µg 118€
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786704 25 µg 231€
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786706 100 µg 685€
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Description

Human cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) is a member of the IgG superfamily. It encodes a 223 amino acid protein and possesses a single extracellular V domain, a transmembrane domain, and a cytoplasmic domain. CTLA-4 is ~30% homologous with CD28, and both bind to the same ligands (B7-1/CD80 and B7-2/CD86) although CTLA-4 binds with higher affinity than CD28. Parallel recognition of a specific MHC-peptide complex by the T-cell receptor and of CD80 or CD86 by the costimulatory receptor CD28 results in T cell activation, cytokine production, proliferation, and differentiation. After the T cells' activation process and upregulation of CTLA-4, co-ligation of the TCR to the MHC-peptide complex and CTLA-4 to its ligands results in cell cycle arrest and ends T cell activation. It has been suggested that the negative regulatory mechanism of CTLA-4 is mediated through trans-endocytosis of CD80 and CD86 (capture of ligands from opposing cells). The ligands are removed from the original cell and degraded inside of CTLA-4-expressing cells. As a result, the action of CD28 is impaired. Alternative splice variants of CTLA-4 have been described in mice and humans that have transcripts that skip exon 2 (ligand-binding domain) or exon 3 (transmembrane domain). The ligand independent CTLA-4 is expressed in mice, but not in humans, and soluble CTLA-4 (sCTLA-4) is expressed in mice and humans. CTLA-4 is constitutively expressed by Tregs, and they are an important source of sCTLA-4. It has been suggested that sCTLA-4 potentiates regulatory T cell function. High levels of sCTLA-4 in Tregs have been associated with type 1 diabetes (T1D) and other autoimmune diseases, such as Grave's disease and myasthenia gravis. Also, CTLA-4 has been linked with proliferation and survival of chronic lymphocytic leukemia. CTLA-4 is a target for therapeutic intervention; in fact, a monoclonal antibody against CTLA-4 (Ipilimumab) has been approved for the treatment of melanoma.

Product Details
Technical Data Sheet (pdf)

Product Details

Source
Human CTLA-4, amino acids Ala31-Phe162 (Accession # BC070162) with a C-terminal human IgG1 Fc tag was expressed in CHO cells.
Molecular Mass
The unlabeled 360 amino acid recombinant protein has a predicted molecular mass of approximately 39.9 kD. The DTT-reduced and non-reduced glycosylated protein migrate at approximately 56 kD and 105 kD respectively by SDS-PAGE. The N-terminal amino acid is Ala.
Purity
>95%, as determined by Coomassie stained SDS-PAGE.
Formulation
0.22 µm filtered protein solution is in PBS, pH 7.2, 5% glycerol.
Endotoxin Level
Less than 0.01 ng/µg cytokine as determined by the LAL method.
Concentration
10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & Handling
Unopened vial can be stored between 2°C and 8°C for one month, at -20°C for six months, or at -70°C for one year. For maximum results, quick spin vial prior to opening. Stock solutions should be prepared at no less than 50 - 100 µg/mL in sterile buffer (PBS, HPBS, DPBS, and EBSS) containing carrier protein such as 0.2 - 1% BSA or HSA and stored in working aliquots at -20°C to -70°C. Avoid repeated freeze/thaw cycles.
Activity
When Recombinant Human B7-2/CD86-Fc Chimera (Cat. No. 775602) is immobilized at 1.0 µg/mL, biotinylated human CTLA-4-Fc Chimera binds with an EC50 of 12 - 48 ng/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding.
Application

Bioassay

Application Notes

BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com.

Antigen Details

Structure
Disulfide-linked homodimer, biotinylated via amines
Distribution

Activated T cells, CD4, CD8, and Tregs

Function
Inhibits T cell activation by reducing IL-2 production and IL-2R expression, also arresting T cells in G1 phase of the cell cycle; protects against the development of autoimmunity
Interaction
Antigen presenting cells, B cells
Ligand/Receptor
CD86, CD80
Bioactivity
Measured by its ability to bind mouse CD200R1
Cell Type
Antigen-presenting cells, B cells, Dendritic cells, Monocytes, T cells
Biology Area
Adaptive Immunity, Immuno-Oncology, Immunology
Molecular Family
Immune Checkpoint Receptors, Soluble Receptors
Antigen References
  1. Linsley PS, et al. 1991. J. Exp. Med. 174:561-9. 
  2. Pentcheva-Hoang T, et al. 2004. Immunity 21:401-13. 
  3. Qureshi OS, et al. 2011. Science 10.1126/science.1202947.
  4. Gerold KD, et al. 2011. Diabetes. 10.2337/db11-0130. 
  5. Rydén A, et al. 2012. Diabetes Metab. Res. Rev. 10.1002/dmrr.1286. 
  6. Mittal AK, et al. 2013. PloS One 8:e70352.
  7. Ward FJ, et al. 2013. Eur. J. Immunol. 10.1002/eji.201242529. 
Gene ID
1493 View all products for this Gene ID
UniProt
View information about CTLA-4 on UniProt.org
Go To Top Version: 1    Revision Date: 09.16.2019

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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