Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Adhesion

Integrin α4 (D2E1) XP® Rabbit mAb #8440

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC F H M R Endogenous 70, 140, 150, Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Integrin α4 (D2E1) XP® Rabbit mAb recognizes endogenous levels of total integrin α4 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser1027 of human integrin α4 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Integrin α4 (D2E1) XP® Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells using Integrin α4 (D2E1) XP® Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of Jurkat (positive; left) and K-562 (negative; right) cells using Integrin α4 (D2E1) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Background

Integrins are α/β heterodimeric cell surface receptors that play a pivotal role in cell adhesion and migration, as well as in growth and survival (1,2). The integrin family contains at least 18 α and 8 β subunits that form 24 known integrins with distinct tissue distribution and overlapping ligand specificities (3). Integrins not only transmit signals to cells in response to the extracellular environment (outside-in signaling), but also sense intracellular cues to alter their interaction with the extracellular environment (inside-out signaling) (1,2).A pair of important α4 integrins, α4β1 and α4β7, interact with VCAM-1, fibronectin, and MAdCAM-1 at cell adhesions (3). Gene knockout and antibody blocking research reveal that α4 integrins play important roles in embryonic liver and heart development and in fetal lymphocyte homing (4-6). Phosphorylation at Ser988 within the cytoplasmic tail of integrin α4 blocks binding to paxillin and promotes leading edge migration (7,8).On SDS-PAGE, integrin α4 can migrate at several different apparent molecular sizes, a 150 kDa mature protein and a 140 kDa precursor protein (a 180 kDa protein also exists under mild non-reducing conditions) (9). Integrin α4 has a cleavage site at Arg558, which results in a small portion of the protein as either an 80 kDa N-terminal or 70 kDa C-terminal fragment (10).

  1. Hood, J.D. and Cheresh, D.A. (2002) Nat Rev Cancer 2, 91-100.
  2. Liu, S. et al. (2000) J Cell Sci 113 ( Pt 20), 3563-71.
  3. Plow, E.F. et al. (2000) J Biol Chem 275, 21785-8.
  4. Bonder, C.S. et al. (2005) Immunity 23, 153-63.
  5. Arroyo, A.G. et al. (1999) Immunity 11, 555-66.
  6. Yang, J.T. et al. (1995) Development 121, 549-60.
  7. Nishiya, N. et al. (2005) Nat Cell Biol 7, 343-52.
  8. Alon, R. et al. (2005) J Cell Biol 171, 1073-84.
  9. Teixidó, J. et al. (1992) J Biol Chem 267, 1786-91.
  10. Pujades, C. et al. (1996) Biochem J 313 ( Pt 3), 899-908.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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