Immunofluorescence analysis of Rhodopsin Kinase 1a was done on 70% confluent log phase MCF-7 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 Rhodopsin Kinase 1a (G8) Mouse Monoclonal Antibody (MA1720) 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 (Product # 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 is a merged image showing Cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Chicken, Human, Mouse, Rat|
|Published species reactivity||Dog, Rat, Bovine, Mouse, Human|
|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 *|
|Immunocytochemistry (ICC)||2-3 µg/ml|
|Immunofluorescence (IF)||2-3 ug/ml|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1 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.
MA1-720 detects G-protein-associated rhodopsin kinase 1a (GRK1a) from human, mouse, rat, bovine and chicken tissues.
MA1-720 has been successfully used in Western blot, immunofluorescence and immunoprecipitation procedures. By Western blot, this antibody detects an ~60 kDa protein representing GRK1a from bovine rod outer segments. Immunofluorescence staining of GRK1a in bovine retina with MA1-720 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 MA1-720 immunogen is full length human GRK1. This antibody recognizes an epitope in the C-terminal region of GRK1a.
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.
The age-regulating protein klotho is vital to sustain retinal function.
MA1-720 was used in western blot to study the expression of klotho in murine retina and its role in maintaining retinal function
|Reish NJ,Maltare A,McKeown AS,Laszczyk AM,Kraft TW,Gross AK,King GD||Investigative ophthalmology and visual science (54:6675)||2013|
Modulation of mouse rod response decay by rhodopsin kinase and recoverin.
MA1-720 was used in western blot to study the role of rhodopsin kinase and recoverin in murine rod response decay
|Chen CK,Woodruff ML,Chen FS,Chen Y,Cilluffo MC,Tranchina D,Fain GL||The Journal of neuroscience : the official journal of the Society for Neuroscience (32:15998)||2012|
Replacing the rod with the cone transducin subunit decreases sensitivity and accelerates response decay.
MA1-720 was used in western blot to investigate the mechanism why cone vision is less sensitive than rod vision
|Chen CK,Woodruff ML,Chen FS,Shim H,Cilluffo MC,Fain GL||The Journal of physiology (588:3231)||2010|
CREB1/ATF1 activation in photoreceptor degeneration and protection.
MA1-720 was used in western blot to study the expression levels of p-CREB1, CREB1, and ATF1 in photoreceptor disease and protection.
|Beltran WA,Allore HG,Johnson E,Towle V,Tao W,Acland GM,Aguirre GD,Zeiss CJ||Investigative ophthalmology and visual science (50:5355)||2009|
Arrestin competition influences the kinetics and variability of the single-photon responses of mammalian rod photoreceptors.
MA1-720 was used in western blot to study the effect of GRK1 and arrestin 1 on the kinetics and variability of single-photon responses of a rod.
|Doan T,Azevedo AW,Hurley JB,Rieke F||The Journal of neuroscience : the official journal of the Society for Neuroscience (29:11867)||2009|
High levels of retinal membrane docosahexaenoic acid increase susceptibility to stress-induced degeneration.
MA1-720 was used in western blot to study the effect of retinal membrane docosahexaenoic acid on stress-induced retinal degeneration in mice.
|Tanito M,Brush RS,Elliott MH,Wicker LD,Henry KR,Anderson RE||Journal of lipid research (50:807)||2009|
Overexpression of rhodopsin alters the structure and photoresponse of rod photoreceptors.
MA1-720 was used in western blot to study how rhodopsin's overexpression affects the structure and photoresponse of rod photoreceptors
|Wen XH,Shen L,Brush RS,Michaud N,Al-Ubaidi MR,Gurevich VV,Hamm HE,Lem J,Dibenedetto E,Anderson RE,Makino CL||Biophysical journal (96:939)||2009|
Transducin activation state controls its light-dependent translocation in rod photoreceptors.
MA1-720 was used in western blot to investigate the role of the GTP-hydrolysis on Gtalpha for light-dependent translocation in rod photoreceptors
|Kerov V,Chen D,Moussaif M,Chen YJ,Chen CK,Artemyev NO||The Journal of biological chemistry (280:41069)||2005|
Functional characterization of mouse RDH11 as a retinol dehydrogenase involved in dark adaptation in vivo.
MA1-720 was used in western blot to demonstrate the effect of mRDH11 disruption on retinoid metabolism in photoreceptor inner segments .
|Kasus-Jacobi A,Ou J,Birch DG,Locke KG,Shelton JM,Richardson JA,Murphy AJ,Valenzuela DM,Yancopoulos GD,Edwards AO||The Journal of biological chemistry (280:20413)||2005|
The bovine iris-ciliary epithelium expresses components of rod phototransduction.
MA1-720 was used in western blot to investigate the level of phototransduction proteins in bovine iris-ciliary epithelium
|Ghosh S,Salvador-Silva M,Coca-Prados M||Neuroscience letters (370:7)||2004|
Leber congenital amaurosis linked to AIPL1: a mouse model reveals destabilization of cGMP phosphodiesterase.
MA1-720 was used in western blot to investigate the essential role of photoreceptor-specific gene Aryl hydrocarbon interacting protein like 1 (AIPL1) in the retina of a mouse Leber congenital amaurosis model.
|Ramamurthy V,Niemi GA,Reh TA,Hurley JB||Proceedings of the National Academy of Sciences of the United States of America (101:13897)||2004|
Phosphorylation of photolyzed rhodopsin is calcium-insensitive in retina permeabilized by alpha-toxin.
MA1-720 was used in western blot to investigate the influence of calcium on rhodopsin phosphorylation in retina
|Otto-Bruc AE,Fariss RN,Van Hooser JP,Palczewski K||Proceedings of the National Academy of Sciences of the United States of America (95:15014)||1998|
Tubby is required for trafficking G protein-coupled receptors to neuronal cilia.
MA1-720 was used in immunohistochemistry to study the role of tubby in neuronal ciliary trafficking of GPCRs
|Sun X,Haley J,Bulgakov OV,Cai X,McGinnis J,Li T||Cilia (1:null)||2012|
Flupirtine attenuates sodium nitroprusside-induced damage to retinal photoreceptors, in situ.
MA1-720 was used in immunohistochemistry to study the mechanism by which flupirtine attenuates sodium nitroprusside-induced damage to retinal photoreceptors
|Fawcett RJ,Osborne NN||Brain research bulletin (73:278)||2007|
Differentiation of embryonic stem cells to retinal cells in vitro.
MA1-720 was used in immunocytochemistry to develop a method for the in vitro differentiation of embryonic stem cells to retinal cells
|Zhao X,Liu J,Ahmad I||Methods in molecular biology (Clifton, N.J.) (330:401)||2006|
Cone deactivation kinetics and GRK1/GRK7 expression in enhanced S cone syndrome caused by mutations in NR2E3.
MA1-720 was used in immunocytochemistry to study the visual abnormalities in patients with NR2E3 mutations
|Cideciyan AV,Jacobson SG,Gupta N,Osawa S,Locke KG,Weiss ER,Wright AF,Birch DG,Milam AH||Investigative ophthalmology and visual science (44:1268)||2003|