Recombinant Mouse Integrin alpha 4 beta 1 Protein, CF

• Reactivity: Mu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Mouse Integrin alpha 4 beta 1 Protein, CF Summary

• Details of Functionality: Measured by the ability of the immobilized protein to support the adhesion of CHO Chinese hamster ovary cells transfected with VCAM-1. When 5 x 10⁴ cells/well are added to Recombinant Mouse Integrin alpha 4 beta 1 coated plates (10 μg/mL, 100 μL/well), approximately 55-85% will adhere after 1 hour at 37 °C.
  Optimal dilutions should be determined by each laboratory for each application.
• Source: Chinese Hamster Ovary cell line, CHO-derived mouse Integrin alpha 4 beta 1 protein
  

  Mouse Integrin alpha 4 Tyr34-Gln970 (Arg591Leu) Accession # AAH68313	GGGSGGGSGGGS	Acidic Tail	6-His tag
  Mouse Integrin beta 1 (Gln2-Asp728) Accession # P09055	GGGSGGGSGGGS	Basic Tail
  N-terminus			C-terminus

• Accession #: AAH68313 (Integrin alpha 4) & P09055 (Integrin beta 1)
• N-terminal Sequence: Tyr34 (Integrin alpha 4) & No results obtained: Gln21 predicted (Integrin beta 1)
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 112.9 kDa (Integrin alpha 4) & 86.4 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: 145-160 kDa and 120-140 kDa, reducing donditions

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: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 200 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 Mouse 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. The 944 aa mouse alpha 4 ECD shares 85% and 94% aa identity with human and rat alpha 4, respectively, while the 708 aa mouse beta 1 ECD shares 98% aa identity with rat and 93-94% aa identity with human, bovine, porcine, ovine, canine and feline beta 1. 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 (5-9). Deletion is lethal in the mouse embryo due to faulty placentation and development of the epicardium and coronary vessels (6, 9). In the adult, alpha 4 beta 1 primarily regulates immune cell migration (10-12). Circulating leukocyte alpha 4 beta 1 is rapidly activated by inflamed endothelial cells that present VCAM-1 and chemokines such as SDF-1 (10). This activation facilitates rolling, firm adhesion, and extravasation. In humans, interfering with leukocyte migration via the therapeutic alpha 4 beta 1antibody Natalizumab can reduce the severity of autoimmune disorders such as multiple sclerosis (11). Natalizumab can also mobilize hematopoietic precursors from the bone marrow by impeding their interaction with stromal cell VCAM-1 (7, 11).  During immune cell activation, alpha 4 beta 1 can function as a costimulatory molecule (12, 13).

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. Eshghi, S. et al. (2007) J. Cell Biol. 177:871.
6. Yang, J.T. et al. (1995) Development 121:549.
7. Sheppard, A.M. et al. (1994) Cell Adhes. Commun. 2:27.
8. Haack, H. and R.O. Hynes (2001) Dev. Biol. 233:38.
9. Grazioli, A. et al. (2006) Dev. Biol. 293:165.
10. Alon, R. and K. Ley (2008) Curr. Opin. Cell Biol. 20:525.
11. Bauer, M. et al. (2009) Proc. Natl. Acad. Sci. USA 106:1920.
12. Banerjee, E.R. et al. (2008) Exp. Hematol. 36:1004.
13. Nguyen, K. et al. (2008) Immunity 28:810.

Publications for Integrin alpha 4 beta 1 (6054-A4)(1)

Publication using 6054-A4

• Azab W, Zajic L, Osterrieder N The role of glycoprotein H of equine herpesviruses 1 and 4 (EHV-1 and EHV-4) in cellular host range and integrin binding. Vet Res, 2012-08-21;43(1):61. 2012-08-21 [PMID: 22909178] (Bioassay, Equine)

Bioinformatics

• Uniprot:
  • AAH68313 (Integrin alpha 4) & P09055 (Integrin beta 1)()