Recombinant Human CD4 Fc Chimera Avi-tag Protein, CF

Catalog #: AVI10965 Datasheet

Biotinylated

Catalog #	Availability	Size / Price	Qty
AVI10965-050

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Recombinant Human CD4 Fc Chimera Avi-tag Protein, CF Summary

Learn more about Avi-tag Biotinylated Proteins

Product Specifications

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 Human CD4 Antibody (Catalog # AF-379-NA) is immobilized at 2.00 μg/mL (100 μL/well), Biotinylated Recombinant Human CD4 Fc Chimera Avi-tag binds with an ED₅₀ of 2.00-12.0 ng/mL.

Source

Chinese Hamster Ovary cell line, CHO-derived human CD4 protein

Human CD4 (Lys26-Trp390) Accession # P01730.1	IEGRMD	Human IgG₁ Fc (Pro100-Lys330)	Avi-tag
N-terminus			C-terminus

Accession #

P01730.1

N-terminal Sequence
Analysis

Lys26

Structure / Form

Disulfide-linked homodimer

Biotinylated via Avi-tag

Predicted Molecular Mass

69 kDa

SDS-PAGE

78-87 kDa, under reducing conditions.

Product Datasheets

AVI10965

Product Datasheet

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.

AVI10965

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.

Scientific Data

Binding Activity

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When Human CD4 Antibody (AF-379-NA) is immobilized at 2.00 μg/mL (100 μL/well), Biotinylated Recombinant Human CD4 Fc Chimera Avi-tag Protein (Catalog # AVI10965) binds with an ED₅₀ of 2.00‑12.0 ng/mL.

SDS-PAGE

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2 μg/lane of Biotinylated Recombinant Human CD4 Fc Chimera Avi-tag Protein (Catalog # AVI10965) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 78-87 kDa and 160-170 kDa, respectively.

Background: CD4

CD4, also known as L3T4, T4, and W3/25, is an approximately 55 kDa type I transmembrane glycoprotein that is expressed predominantly on thymocytes and a subset of mature T lymphocytes. It is a standard phenotype marker for the identification of T cell populations (1). Mature human CD4 consists of a 371 amino acid (aa) extracellular region containing four immunoglobulin-like domains, a 22 aa transmembrane segment, and a 40 aa cytoplasmic domain (2). Within the ECD, human CD4 shares approximately 52% aa sequence identity with mouse and rat CD4. CD4 is expressed along with CD8 on double positive T cells during their development in the thymus. Either CD4 or CD8 expression is then lost, giving rise to single positive (SP) CD4+ or CD8+ mature T cells (3). CD4+ SP cells, also known as T helper cells, further differentiate into multiple subsets of CD4+ cells including Th1, Th2, Th17, Tfh, and Treg cells which regulate humoral and cellular immunity (4). CD4 is reexpressed on circulating CD8+ T cells upon activation and contributes to their cytotoxic effector activity (5). In human, CD4 is additionally expressed on macrophages, neutrophils, monocytes, NK cells, and neurons and glial cells in the brain (6-9). Similar CD4 distribution between species cannot be assumed as demonstrated by its presence on macrophages in human and rat but not in mouse (6). CD4 binds directly to MHC class II molecules on antigen presenting cells (10). This interaction contributes to the formation of the immunological synapse which is focused around the TCR-MHC class II-antigenic peptide interaction (1, 11). Palmitoylation of two cysteine residues in the cytoplasmic tail of CD4 promotes the localization of CD4 in lipid rafts and its ability to augment TCR signaling via activation of the tyrosine kinase Lck (12). CD4 also functions as a chemotactic receptor for IL-16 and, in human, as a co-receptor for the gp120 surface glycoprotein of HIV-1 (7, 13-15). Our Avi-tag Biotinylated human CD4 Fc Chimera features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

References

Vignali, D.A.A. (2010) J. Immunol. 184:5933.
Maddon, P.J. et al. (1985) Cell 42:93.
Alarcon, B. and H.M. van Santen (2010) Sci. Signal. 3:pe11.
Wan, Y.Y. and R.A. Flavell (2009) Mol. Cell Biol. 1:20.
Kitchen, S.G. et al. (2005) Proc. Natl. Acad. Sci. 102:3794.
Crocker, P.R. et al. (1987) J. Exp. Med. 166:613.
Biswas, P. et al. (2003) Blood 101:4452.
Bernstein, H.B. et al. (2006) J. Immunol. 177:3669.
Funke, I. et al. (1987) J. Exp. Med. 165:1230.
Doyle, C. and J.L. Strominger (1987) Nature 330:256.
Huppa, J.B. et al. (2010) Nature 463:963.
Fragoso, R. et al. (2003) J. Immunol. 170:913.
Cruikshank, W.W. et al. (1994) Proc. Natl. Acad. Sci. 91:5109.
Klatzmann, D. et al. (1984) Nature 312:767.
Dagleish, A.G. et al. (1984) Nature 312:763.

Entrez Gene IDs

920 (Human); 12504 (Mouse); 24932 (Rat); 403931 (Canine); 101864991 (Cynomolgus Monkey); 493775 (Feline)

Alternate Names

CD_antigen: CD4; CD4 antigen (p55); CD4 antigen; CD4 molecule; CD4 receptor; CD4; CD4mut; T-cell surface antigen T4/Leu-3; T-cell surface glycoprotein CD4