Immunofluorescent analysis of Rhodopsin Kinase 1a/b (green) in murine cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes at room temperature and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Rhodopsin Kinase 1a/b Monclonal Antibody (D11) (Product # MA1-721 ) at a dilution of 1:200 and incubated overnight in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Images were taken at a 60X magnification.
|Tested species reactivity||Bovine, Human, Mouse|
|Published species reactivity||Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Full length human GRK1.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:1,000|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1,000|
* 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.
MA1-721 detects G-protein-associated rhodopsin kinase 1a/b (GRK1a/b) from human and bovine tissues. This antibody does not cross-react with other G protein-coupled kinases.
MA1-721 has been successfully used in Western blot, immunofluorescence, and immunoprecipitation experiments. By Western blot, this antibody detects an ~60 kDa protein representing GRK 1a/b. MA1-721 detects both in vitro expressed splice variants by Western blot but was not able to detect significant amounts of GRK1b from human retina. Immunofluorescence staining of GRK1a/b in bovine retina with MA1-721 results in intense staining primarily in cone outer segments, but also in rod outer segments. Weak staining is observed in somata and synaptic terminals of cones and inner segments of rods.
The MA1-721 immunogen is full length human GRK1a. This antibody recognizes an N-terminal epitope of human GRK1 which is conserved in both GRK1a and GRK1b.
The sensation of sight is the result of a cascade of events starting with the interaction of photoactivated rhodopsin with a protein called transducin. Rhodopsin kinase is a G-protein-coupled Ser/Thr kinase, also known as GRK1, which is a key element in the regulation of this cascade. Following phosphorylation by GRK1, arrestin is recruited to phospho-rhodopsin quenching its phototransductive activity by preventing further interaction with transducin. By breaking the cycle of phototransduction, GRK1 plays an important role in the restoration of the system for subsequent visual events.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
GAP-independent termination of photoreceptor light response by excess gamma subunit of the cGMP-phosphodiesterase.
MA1-721 was used in western blot to study the effect of the cGMP-phosphodiesterase gamma subunit on photoreceptor light response termination.
|Tsang SH,Woodruff ML,Chen CK,Yamashita CY,Cilluffo MC,Rao AL,Farber DB,Fain GL||The Journal of neuroscience : the official journal of the Society for Neuroscience (26:4472)||2006|
A short, highly active photoreceptor-specific enhancer/promoter region upstream of the human rhodopsin kinase gene.
MA1-721 was used in immunohistochemistry to define a photoreceptor-specific enhancer-promoter upstream of the Rk gene.
|Young JE,Vogt T,Gross KW,Khani SC||Investigative ophthalmology and visual science (44:4076)||2003|
GRK1-dependent phosphorylation of S and M opsins and their binding to cone arrestin during cone phototransduction in the mouse retina.
MA1-721 was used in immunohistochemistry to investigate the role of GRK1-dependent phosphorylation of cone opsins during the murine cone phototransduction.
|Zhu X,Brown B,Li A,Mears AJ,Swaroop A,Craft CM||The Journal of neuroscience : the official journal of the Society for Neuroscience (23:6152)||2003|