Technical Data

C2255-80D
Clone Type
Monoclonal
Host
Mouse
Source
Rat
Isotype
IgG2b
Clone Number
12K25
Grade
Affinity Purified
Applications
WB
Crossreactivity
Rt
Shipping Temp
Blue Ice
Storage Temp
-20°C
Mouse Anti-CD4 (T-cell Surface Glycoprotein CD4, T-cell Surface Antigen T4/Leu-3, W3/25 Antigen)

CD4, also known as L3T4, T4, and W3/25, is an ~55kD 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. Mature rat CD4 consists of a 367aa extracellular region containing four immunoglobulin-like domains, a 23aa transmembrane segment, and a 40aa cytoplasmic domain. 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. 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. CD4 is reexpressed on circulating CD8+ T cells upon activation and contributes to their cytotoxic effector activity. In the rat, CD4 is additionally expressed on macrophages and selected subsets of dendritic cells. Similar CD4 distribution between species cannot be assumed as demonstrated by its presence on macrophages in human and rat but not in mouse. CD4 binds directly to MHC class II molecules on antigen presenting cells. This interaction contributes to the formation of the immunological synapse which is focused around the TCRMHC class IIantigenic peptide interaction. 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. CD4 also functions as a chemotactic receptor for IL16 and, in human, as a coreceptor for the 120aa surface glycoprotein of HIV1(1114).CD4 associates with CD44 on the cell surface to potentiate CD44 mediated cell adhesion.

Applications
Suitable for use in Western Blot Other applications not tested.
Recommended Dilution
Western Blot: 0.2ug/ml Optimal dilutions to be determined by the researcher.
Storage and Stability
Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Immunogen
Recombinant corresponding to aa28-394 from rat CD4 expressed in NSO cell line.
Form
Supplied as a lyophilized powder from PBS, 5% trehalose. Reconstitute with 200ul sterile PBS.
Purity
Purified by Protein A/G affinity chromatography.
Specificity
Recognizes rat CD4. No crossreactivity with canine, cotton rat, feline, human or mouse CD4.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1. Vignali, D.A.A. (2010) J. Immunol. 184:5933. 2. Clark, S.J. et al. (1987) Proc. Natl. Acad. Sci. 84:1649. 3. Alarcon, B. and H.M. van Santen (2010) Sci. Signal. 3:pe11. 4. Wan, Y.Y. and R.A. Flavell (2009) Mol. Cell Biol. 1:20. 5. Kitchen, S.G. et al. (2005) Proc. Natl. Acad. Sci. 102:3794. 6. Crocker, P.R. et al. (1987) J. Exp. Med. 166:613. 7. Liu, L. et al. (1998) J. Immunol. 161:1146. 8. Doyle, C. and J.L. Strominger (1987) Nature 330:256. 9. Huppa, J.B. et al. (2010) Nature 463:963. 10. Fragoso, R. et al. (2003) J. Immunol. 170:913. 11. Cruikshank, W.W. et al. (1994) Proc. Natl. Acad. Sci. 91:5109. 12. Klatzmann, D. et al. (1984) Nature 312:767. 13. Dagleish, A.G. et al. (1984) Nature 312:763. 14. Biswas, P. et al. (2003) Blood 101:4452. 15. Umberto D. et al. (1999) International Immunology 11:1085.
USBio References
No references available
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