Recombinant Cynomolgus Integrin alpha 4 beta 1 Protein, CF

When Recombinant Human VCAM-1/CD106 Fc Chimera (862-VC) is immobilized at 1 µg/mL (100 µL/well), Recombinant Cynomolgus Monkey Integrin alpha 4 beta 1 (Catalog # 10527-A4) binds with an ED50 of 4-40 ng/mL.

• Reactivity: Pm-Cm
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Cynomolgus Integrin alpha 4 beta 1 Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human VCAM-1/CD106 Fc Chimera (Catalog # 862-VC) is immobilized at 1 µg/mL (100 µL/well), Recombinant Cynomolgus Monkey Integrin alpha 4 beta 1 (Catalog # 10527-A4) binds with an ED₅₀ of 4-40 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived cynomolgus monkey Integrin alpha 4 beta 1 protein
  

  Cynomolgus Monkey Integrin alpha 4 (Tyr34-Gln970) Accession # XP_005573683.1	HP + 2x GGGSGGGS	Acidic Tail	6-His tag

  Cynomolgus Monkey Integrin beta 1 (Gln21-Asp728) Accession # XP_005564991.1	HP + 2x GGGSGGGS	Basic Tail	HA-tag

  N-terminus C-terminus

• Accession #: XP_005573683.1 (Integrin alpha 4) & XP_005564991.1
• N-terminal Sequence: Tyr34 (Integhrin alpha 4) & Gln21 inferred from enzymatic pyroglutamate treatment revealing Thr22 (Integrin beta 1)
• Protein/Peptide Type: Recombinant Proteins
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 113 kDa (Integrin alpha 4) & 88 kDa (Integrin beta 1).
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 120-160 kDa, under reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 250 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Cynomolgus Integrin alpha 4 beta 1 Protein, CF

• CD49d
• IA4
• Integrin alpha 4 beta 1

Background

Integrin alpha 4 beta 1, also called VLA4, is an integrin family adhesion receptor that shares the beta 1 subunit with eleven other family members and the alpha 4 subunit with integrin alpha 4 beta 7 (1-4). The non-covalent heterodimer of 150 kDa alpha 4/CD49d and 130 kDa beta 1/CD29 type I transmembrane glycoprotein subunits mediates cell adhesion to VCAM-1/CD106 on other cells and the CS-1 fragment of fibronectin in the extracellular matrix (2-4). The alpha 4 extracellular domain (ECD) contains an N-terminal beta -propeller structure, followed by domains termed thigh, calf-1 and calf-2 (1). The beta 1 ECD contains a vWFA domain, which interacts with the alpha 4 beta -propeller to form a binding domain when the dimer is in active, extended and open conformation. Each subunit has a transmembrane sequence and a short cytoplasmic tail. Within the ECD, cynomolgus alpha 4 shares 97% sequence idently with human alpha 4, while cynomolgus beta 1 shares 99.9% sequence identity with human beta 1 ECD. Five alternate splice forms of the human beta 1 cytoplasmic domain, including one antagonistic form, vary by 12 to 48 aa and show differential expression patterns (5). Leukocytes (except for neutrophils), erythroid precursors and some non-hematopoietic cells such as epicardial, endothelial and smooth muscle precursors, Schwann cells, and chorionic cells express alpha 4 beta 1 (6-10). Deletion is lethal in the mouse embryo due to faulty placentation and development of the epicardium and coronary vessels (7, 10). In the adult, alpha 4 beta 1 primarily regulates immune cell migration (11-13). Circulating leukocyte alpha 4 beta 1 is rapidly activated by inflamed endothelial cells that present VCAM-1 and chemokines such as SDF-1 (11). This activation facilitates rolling, firm adhesion, and extravasation. Interfering with leukocyte migration via the therapeutic alpha 4 beta 1antibody Natalizumab can reduce the severity of autoimmune disorders such as multiple sclerosis (12). Natalizumab can also mobilize hematopoietic precursors from the bone marrow by impeding their interaction with stromal cell VCAM-1 (8, 12). During immune cell activation, alpha 4 beta 1 can function as a costimulatory molecule (13, 14).

1. Luo, B-H. et al. (2007) Annu. Rev. Immunol. 25:619.
2. Takada, Y. et al. (1989) EMBO J. 8:1361.
3. Argraves, W.S. et al. (1987) J. Cell Biol. 1025:1183.
4. Takada, Y. et al. (2007) Genome Biol. 8:215.
5. Svineng, G. et al. (1998) Biochem. J. 330:1255.
6. Eshghi, S. et al. (2007) J. Cell Biol. 177:871.
7. Yang, J.T. et al. (1995) Development 121:549.
8. Sheppard, A.M. et al. (1994) Cell Adhes. Commun. 2:27.
9. Haack, H. and R.O. Hynes (2001) Dev. Biol. 233:38.
10. Grazioli, A. et al. (2006) Dev. Biol. 293:165.
11. Alon, R. and K. Ley (2008) Curr. Opin. Cell Biol. 20:525.
12. Bauer, M. et al. (2009) Proc. Natl. Acad. Sci. USA 106:1920.
13. Banerjee, E.R. et al. (2008) Exp. Hematol. 36:1004.
14. Nguyen, K. et al. (2008) Immunity 28:810.

Bioinformatics

• Uniprot:
  • XP_005573683.1 (Integrin alpha 4) & XP_005564991.1()