Rat CD4 Antibody

Catalog #: AF6439 Datasheet

Catalog #	Availability	Size / Price	Qty
AF6439
AF6439-SP

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Rat CD4 Antibody Summary

Species Reactivity

Rat

Specificity

Detects rat CD4 in direct ELISAs and Western blots. In direct ELISAs, approximately 3% cross-reacitvity with recombinant feline CD4 is observed, and less than 1% cross-reactivity with recombinant human CD4 and recombinant canine CD4 is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant rat CD4
Lys28-Thr394
Accession # P05540

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Flow Cytometry

2.5 µg/10⁶ cells

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Rat CD4 by Western Blot. Western blot shows lysates of rat CD4⁺T cells and rat spleen tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Rat CD4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6439) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for CD4 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Flow Cytometry

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Detection of rat CD4 in Rat Splenocytes by Flow Cytometry. Rat splenocytes were stained with Sheep Anti-Rat CD4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6439, filled histogram) or control antibody (Catalog # 5-001-A, open histogram), followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127).

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

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 rat CD4 consists of a 367 amino acid (aa) extracellular region containing four immunoglobulin-like domains, a 23 aa transmembrane segment, and a 40 aa cytoplasmic domain (2). Within the ECD, rat CD4 shares 52% and 73% aa sequence identity with human and mouse CD4, respectively. 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 re‑expressed on circulating CD8⁺ T cells upon activation and contributes to their cytotoxic effector activity (5). In the rat, CD4 is additionally expressed on macrophages and selected subsets of dendritic cells (6, 7). 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 (8). This interaction contributes to the formation of the immunological synapse which is focused around the TCR-MHC class II-antigenic peptide interaction (1, 9). 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 (10). 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 (11‑14). CD4 associates with CD44 on the cell surface to potentiate CD44-mediated cell adhesion (15).

References

Vignali, D.A.A. (2010) J. Immunol. 184:5933.
Clark, S.J. et al. (1987) Proc. Natl. Acad. Sci. 84:1649.
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.
Liu, L. et al. (1998) J. Immunol. 161:1146.
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.
Biswas, P. et al. (2003) Blood 101:4452.
Umberto D. et al. (1999) International Immunology 11:1085.

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