Western blot analysis of phosphorylated GIT1 was performed by loading human umbilical vein endothelial cell lysates with and without shear stress (at 10 dyn/cm^2) per well onto an SDS PAGE gel. Proteins were transferred to a membrane and blocked with SuperBlock T20 (TBS) Blocking Buffer. The membrane was probed with a Phospho-GIT1 pTyr554 Antibody (Product # PA5-12982) at a dilution of 1:200, washed, and probed with a HRP-conjugated secondary antibody for chemiluminescent detection. β-actin detection (lower panel) was used as loading control. Data courtesy of the Innovators Program.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||KLH conjugated synthetic phosphopeptide corresponding to amino acid residues surrounding Y554 of human GIT1|
|Purification||Antigen affinity chromatography|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Dot blot (DB)||1:500|
|Western Blot (WB)||1:200|
* 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.
This antibody is predicted to react with mouse and rat based on sequence homology.
PA5-12982 was used in western blot to show induction of phophorylation of GIT1 on Y554 in lysates of human umbilical vein endothelial cells.
GIT1 is a GTPase-activating protein for the ADP ribosylation factor family. It may serve as a scaffold to bring together molecules to form signaling modules controlling vesicle trafficking, adhesion and cytoskeletal organization, increases the speed of cell migration, as well as the size and rate of formation of protrusions, possibly by targeting PAK1 to adhesions and the leading edge of lamellipodia. It sequesters inactive non-tyrosine-phosphorylated paxillin in cytoplasmic complexes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.