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Recombinant Human Integrin alpha L beta 2 Protein, CF

Catalog # 3868-AV | R&D Systems, Inc. a Bio-Techne Brand
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3868-AV-050

Key Product Details

Source

CHO

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Integrin alpha L beta 2 protein
Human Integrin alphaL
(Tyr26-Met1089)
Accession # P20701
acidic tail
Human Integrin beta2
(Gln23-Asn700)
Accession # P05107
basic tail
N-terminus C-terminus

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Tyr26 (Integrin alphaL) & No results obtained: Gln23 predicted (Integrin beta2)

Predicted Molecular Mass

121.7 kDa (Integrin alphaL), 79 kDa (Integrin beta2)

SDS-PAGE

160 kDa and 100 kDa, reducing conditions

Activity

Measured by the ability of the immobilized protein to support the adhesion of CHO Chinese hamster ovary cells transfected with ICAM-1.
When 5 x 104 cells/well are added to Integrin alphaL beta2-coated plates (5 µg/mL, 100 µL/well), 60‑80% of the cells will adhere after 1 hour at 37° C in the presence of 1 mM MnCl2.

Formulation, Preparation and Storage

3868-AV
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution
Reconstitute at 100 μg/mL in sterile PBS.

Reconstitution Buffer Available:
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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.

Background: Integrin alpha L beta 2

Integrin alphaL beta2, also called LFA-1, is one of three beta2 integrin adhesion proteins. The non-covalent heterodimer of 180 kDa alphaL/CD11a and 95 kDa beta2/CD18 integrin subunits is expressed on virtually all leukocytes (1-3). The ligand binding site of LFA-1 is in the N-terminal head region, formed by an interaction of the vWFA (I, I‑like) domains from each subunit, and the alphaL beta-propeller structure (3-5). The alphaL subunit contains domains termed thigh, calf-1 and calf-2, while the beta2 subunit contains a PSI (plexin-semaphorin-integrin) region and four cysteine-rich I-EGF folds (5, 6). Each subunit has a transmembrane sequence and a short cytoplasmic tail connected to the cytoskeleton. Upon activation by “inside-out” signaling, clustering, or Mg2+ or Mn2+ binding to metal ion-dependent adhesion sites (MIDAS) within the vWFA domains, the molecule unfolds from its inactive, “closed” conformation to expose ligand binding sites (3, 6-9). Active alphaL beta2 binds ICAM-1/CD54, ICAM-2, ICAM‑3 and JAM‑A (1, 10-12). The adhesion stabilizes interactions between T cells and antigen-presenting cells, decreases the T cell activation threshold, and facilitates leukocyte transendothelial migration to sites of inflammation (12-14). A constitutively active construct severely impairs immune responses, demonstrating that both activation and de-activation are important (14). Mutations of beta2, especially in the vWFA domain, cause leukocyte adhesion deficiency (LAD-1) and susceptibility to bacterial infections (15). The 1088 amino acid (aa) human alphaL/CD11b ECD is 73-74% aa identical to mouse and rat, and 79-80% aa identical to bovine, porcine, ovine, goat and canine  alphaL ECD. A second human alphaL isoform has 53 aa inserted after aa 954 in the ECD. The 678 aa human beta2/CD18 ECD shares 81-83% aa sequence identity with mouse, rat, bovine, canine, goat, ovine, and porcine beta2 ECD.

References

  1. Larson, R.S. et al. (1989) J. Cell Biol. 108:703.
  2. Kishimoto, T.K. et al. (1987) Cell 48:681.
  3. Hynes, R.O. (2002) Cell 110:673.
  4. Qu, A. and D.J. Leavy (1995) Proc. Natl. Acad. Sci. USA 92:10277.
  5. Shi, M. et al. (2005) J. Biol. Chem. 280:30586.
  6. Beglova, N. et al. (2002) Nat. Struct. Biol. 9:282.
  7. Shimaoka, M. et al. (2003) Immunity 19:391.
  8. Lu, C. et al. (2004) J. Immunol. 173:3972.
  9. Cairo, C.W. et al. (2006) Immunity 25:297.
  10. Nortamo, P. et al. (1991) J. Immunol. 146:2530.
  11. de Fougerolles, A.R. and T.A. Springer (1992) J. Exp. Med. 175:185.
  12. Ostermann, G. et al. (2002) Nat. Immunol. 3:151.
  13. Heit, B. et al. (2005) J. Cell Sci. 118:5205.
  14. Semmrich, M. et al. (2005) J. Exp. Med. 201:1987.
  15. Kishimoto, T.K. et al. (1987) Cell 50:193.

Entrez Gene IDs

3683 (Human)

Gene Symbol

ITGAL

Product Documents for Recombinant Human Integrin alpha L beta 2 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human Integrin alpha L beta 2 Protein, CF

For research use only

Citations for Recombinant Human Integrin alpha L beta 2 Protein, CF (5)

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FAQs for Recombinant Human Integrin alpha L beta 2 Protein, CF

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  • Q: What is the amino acid sequence of the acidic and basic tails?

    A: Acidic and basic tails are added to the protein to help facilitate optimal activity. While we generally include sequence information on the product datasheet, the sequences of these tails are considered confidential information.

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