Recombinant Human CTLA-4 Fc Chimera Avi-tag Protein, CF

Biotinylated
Catalog # Availability Size / Price Qty
AVI7268-050
Recombinant Human CTLA-4 Fc Chimera Avi-tag Protein, CF Binding Activity
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Recombinant Human CTLA-4 Fc Chimera Avi-tag Protein, CF Summary

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its binding ability in a functional ELISA. When Recombinant Human B7‑1/CD80Fc Chimera (Catalog # 10133-B1) is immobilized at 0.5 µg/mL (100 µL/well), Biotinylated Recombinant Human CTLA-4 Fc Chimera Avi-tag (Catalog # AVI7268) binds with an ED50 of 0.4-2.4 ng/mL.
Source
Human embryonic kidney cell, HEK293-derived human CTLA-4 protein
Human CTLA-4
(Ala37-Phe162)
Accession # P16410-1
IEGRMDHuman IgG1
(Pro100-Lys330)
Avi-tag
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
Ala37
Structure / Form
Disulfide-linked homodimer, biotinylated via Avi-tag
Predicted Molecular Mass
42 kDa
SDS-PAGE
53-61 kDa, under reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AVI7268

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Data Images

Binding Activity Binding Activity View Larger

When Recombinant Human B7-1/CD80 Fc Chimera (Catalog # 10133-B1) is immobilized at 0.5 µg/mL, Biotinylated Recombinant Human CTLA-4 Fc Chimera Avi-tag (Catalog # AVI7268) binds with anED50 of 0.4-2.4 ng/mL.

SDS-PAGE SDS-PAGE View Larger

2 μg/lane of Biotinylated Recombinant Human CTLA-4 Fc Chimera Avi-tag (Catalog # AVI7268) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 53-61 kDa and 110-120kDa, respectively.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CTLA-4

CTLA-4 (cytotoxic T-lymphocyte associated protein‑4, designated CD152), is a type I transmembrane T cell inhibitory molecule that is a member of the Ig superfamily (1, 2). Human or mouse CTLA-4 cDNA encodes 223 amino acids (aa) including a 35 aa signal sequence, a 126 aa extracellular domain (ECD) with one Ig-like V-type domain, a 21 aa transmembrane (TM) sequence, and a 41 aa cytoplasmic sequence. It is found as a covalent homodimer of 41-43 kDa (2) Within the ECD, human CTLA-4 shares 68%, 71% and 83‑86% aa sequence identity with mouse, rat and porcine/bovine/rabbit/feline/canine CTLA-4, respectively. A 174 aa form that lacks TM and cytoplasmic sequences (sCTLA-4) is possibly secreted (3-5). Isoforms of 56-79 aa that mainly contain parts of the cytoplasmic domain are reported. In mouse, an isoform lacking the Ig-like domain has ligand-independent inhibitory activity and is termed liCTLA-4 (6). CD28, which is structurally related to CTLA-4, is constitutively expressed on naïve T cells and promotes T cell activation when engaged by B7-2 on antigen-presenting cells (APC) within the immunological synapse (IS) (1, 7, 8). In contrast, CTLA-4 is recruited from intracellular vesicles to the IS beginning 1-2 days after T cell activation (2, 7, 8). It forms a linear lattice with B7-1 on APC, inducing negative regulatory signals and ending T cell activation (9). Abatacept, a therapeutic human CTLA-4-Ig fusion protein (trade name Orencia), competes with CD28 for B7-1 and B7-2 binding and has been used to antagonize T cell activation in autoimmune conditions and to enhance transplant survival (10). Mice deleted for CTLA-4 show no abnormalities until after birth, but then develop lethal autoimmune reactions due to continued T cell activation and poor control by regulatory T cells, which constitutively express CTLA-4 in wild-type mice and humans (11-13).

References
  1. Harper, K. et al. (1991) J. Immunol. 147:1037.
  2. Teft, W.A. et al. (2006) Annu. Rev. Immunol. 24:65.
  3. Magistrelli, G. et al. (1999) Eur. J. Immunol. 29:3596.
  4. Tector, M. et al. (2009) BMC Immunol. 10:51.
  5. Oaks, M.K. and K.M. Hallett (2000) J. Immunol. 164:5015.
  6. Vijayakrishnan, L. et al. (2004) Immunity 20:563.
  7. Pentcheva-Hoang, T. et al. (2004) Immunity 21:401.
  8. Jansson, A. et al. (2005) J. Immunol 175:1575.
  9. Darlington, P.J. et al. (2005) J. Immunol. 175:996.
  10. Platt, A.M. et al. (2010) J. Immunol. 185:1558.
  11. Wing, K. et al. (2008) Science 322:271.
  12. Friedline, R.H. et al. (2009) J. Exp. Med. 206:421.
  13. Jain, N. et al. (2010) Proc. Natl. Acad. Sci. USA 107:1524.
Long Name
Cytotoxic T-lymphocyte-associated Molecule 4
Entrez Gene IDs
1493 (Human); 12477 (Mouse); 403696 (Canine); 102115124 (Cynomolgus Monkey)
Alternate Names
CD; CD152 antigen; CD152; CD152IDDM12; CD28; celiac disease 3; CELIAC3; CTLA4; CTLA-4; cytotoxic T-lymphocyte antigen 4; cytotoxic T-lymphocyte protein 4; Cytotoxic T-lymphocyte-associated antigen 4; cytotoxic T-lymphocyte-associated protein 4; cytotoxic T-lymphocyte-associated serine esterase-4; GRD4; GSE; ICOS; ligand and transmembrane spliced cytotoxic T lymphocyte associated antigen 4

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