Immunofluorescence analysis of Phospho-Integrin beta-1 pThr788/pTyr789 was performed using 90% confluent THP-1 cells treated with 100ng/mL of IFN-gamma for 15 minutes. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-Integrin beta-1 pThr788 /pTyr789 Rabbit Polyclonal Antibody (44-872G) at 2 µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization in the membrane. Panel e is untreated cell with less signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Cat, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human integrin b1 receptor that contains threonines 788 and 789 (based on Swiss Protein database, accession number P05558). The sequence is conserved in human, mouse, rat and chicken.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
CD29 (beta1 integrin subunit, GPIIa) forms non-covalently linked heterodimers with at least 6 different alpha chains (alpha1-alpha6, CD49a-f) determining the binding properties of beta1 (VLA) integrins. These integrins mediate cell adhesion to collagen, fibronectin, laminin and other extracellular matrix (ECM) components. This interaction hinders cell death, whereas disruption of anchorage to ECM leads to apoptosis. Decreased expression of most beta1 integrins correlates with acquiring multidrug resistance of tumour cells during selection in presence of antitumour drug. In platelets, translocation of intracellular pool of beta1 integrins to the plasma membrane following thrombin stimulation. These integrins are also up-regulated in leukocytes during emigration and extravascular migration and appear to be critically involved in regulating the immune cell trafficking from blood to tissue, as well as in regulating tissue damage and disease symptoms related to inflammatory bowel disease. Through a beta1 integrin-dependent mechanism, fibronectin and type I collagen enhance cytokine secretion of human airway smooth muscle in response to IL-1beta.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Mouse||Not Cited||Cellular integrins function as entry receptors for human cytomegalovirus via a highly conserved disintegrin-like domain.||Feire AL,Koss H,Compton T||Proceedings of the National Academy of Sciences of the United States of America (101:15470)||2004|
Focal complex formation in adult cardiomyocytes is accompanied by the activation of beta3 integrin and c-Src.
44-872G was used in western blot to investigate the involvement of beta3 integrin and c-Src in focal adhesion complex formation in adult cardiomyocytes.
|Willey CD,Balasubramanian S,Rodríguez Rosas MC,Ross RS,Kuppuswamy D||Journal of molecular and cellular cardiology (35:671)||2003|