Immunofluorescence analysis of GRK2 was performed using 70% confluent log phase PC-3 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 GRK2 (5D5) Mouse Monoclonal Antibody (MA12013) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) 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 cytoplasmic and membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse|
|Immunogen||Recombinant beta adrenergic receptor kinase 1.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||3-5 ug/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.
MA1-2013 detects the beta andrenergic receptor kinase 1 in human, mouse and rat samples.
MA1-2013 has been successfully used in Western blot, immunocytochemistry, immunofluoresence, immunoprecipitation, and ELISA procedures. By Western blot this antibody detects a ~30 kDa protein representing the beta andrenergeic receptor kinase 1 in human HeLa cells. In immunocytochemistry procedures MA1-2013 recognizes the beta andrenergic receptor kinase 1 in HeLa, Hek-293 and 3T3 cells. Immunofluoresence in HeLa cells yeilds predominantely cytoplasmic membrane associated staining with some staining in the ruffles.
The MA1-2013 immunogen is recombinant beta adrenergic receptor kinase 1.
Beta-adrenergic receptor kinase 1 (BARK1) phosphorylates the G-protein coupled b-adrenergic receptor. The b-adrenergic receptor is the primary target of epinephrine in cardiac muscle and other tissues. Elevated levels of BARK1 have been found in failing human heart tissue along with decreased activity of BAR. Knockout studies show that inhibition of BARK1 in Csq mice with severe cardiomyopathy markedly extended lifespan and improve cardiac health. This protein has also been referred to as G Protein Coupled Receptor Kinase 2.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.