Immunofluorescence analysis of Phospho integrin beta 3 was performed using 70% confluent log phase THP-1 cells. 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-3 pTyr773 Rabbit Polyclonal Antibody (44-876G) at 2µg/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 conjµgate (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 membranous localization. Panel e 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, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human Integrin b3 receptor that contains tyrosine 773 (based on Swiss Protein database, accession number P05106). The sequence is conserved in human, mouse and rat.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|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)||Assay Dependent|
* 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.
The ITGB3 protein product is the integrin beta chain beta 3. Integrins are integral cell-surface proteins composed of an alpha chain and a beta chain. A given chain may combine with multiple partners resulting in different integrins. Integrin beta 3 is found along with the alpha IIb chain in platelets. Integrins are known to participate in cell adhesion as well as cell-surface mediated signalling.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Antithrombotic effects of targeting alphaIIbbeta3 signaling in platelets.
44-876G was used in western blot to characterize mice that lack the 3 C-terminal beta3 residues needed for alphaIIbbeta3 interaction with c-Src, but retain the beta3 residues needed for talin-dependent fibrinogen binding
|Ablooglu AJ,Kang J,Petrich BG,Ginsberg MH,Shattil SJ||Blood (113:3585)||2009|
|Human||Not Cited||Eph kinases and ephrins support thrombus growth and stability by regulating integrin outside-in signaling in platelets.||Prévost N,Woulfe DS,Jiang H,Stalker TJ,Marchese P,Ruggeri ZM,Brass LF||Proceedings of the National Academy of Sciences of the United States of America (102:9820)||2005|
|Cat||Not Cited||Focal complex formation in adult cardiomyocytes is accompanied by the activation of beta3 integrin and c-Src.||Willey CD,Balasubramanian S,Rodríguez Rosas MC,Ross RS,Kuppuswamy D||Journal of molecular and cellular cardiology (35:671)||2003|
||Eph kinases and ephrins support thrombus growth and stability by regulating integrin outside-in signaling in platelets.||Prévost N,Woulfe DS,Jiang H,Stalker TJ,Marchese P,Ruggeri ZM,Brass LF||Proceedings of the National Academy of Sciences of the United States of America (102:9820)||2005|