Immunofluorescent analysis of Rhodopsin (green) showing staining in human retinal tissue (right) compared to a negative control without primary antibody (left). Formalin-fixed tissue was permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Tissue was probed with a Rhodopsin monoclonal antibody (Product # MA1-722) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4°C in a humidified chamber. Tissue was washed with PBST and incubated with a DyLight-488 conjugated secondary antibody in PBS at room temperature in the dark. Nuclei were stained with DAPI (blue) and images were taken at a magnification of 20x.
|Tested species reactivity||Bovine, Human, Mouse, Rat|
|Published species reactivity||Rat, Bovine, Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:1000|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:100-1:1000|
* 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.
MA1-722 detects rhodopsin from human, mouse and bovine retinal samples. Clone 1D4 has also been used successfully in Zebrafish. It does not recognize rhodopsin in zebrafish rods, but instead labels long double cone outer segments that express red opsin (Yin, J. et al, 2012).
MA1-722 has been successfully used in Western blot, IHC (P), immunocytochemistry and immunoprecipitation procedures. By Western blot, this antibody detects an ~40 kDa protein representing rhodopsin from Sf9 cells expressing the bovine gene. Immunocytochemical staining of rhodopsin in human retinal samples results in staining of both rod and cone outer segments.
The MA1-722 immunogen is bovine rhodopsin. The epitope for this antibody has been localized to the C-terminal nine amino acids of bovine rhodopsin known as the 1D4 epitope.
Vision involves the conversion of light into electrochemical signals that are processed by the retina and subsequently sent to and interpreted by the brain. The process of converting light to an electrochemical signal begins when the membrane-bound protein, rhodopsin, absorbs light within the retina. Photoexcitation of rhodopsin causes the cytoplasmic surface of the protein to become catalytically active. In the active state, rhodopsin activates transducin, a GTP binding protein. Once activated, transducin promotes the hydrolysis of cGMP by phosphodiesterase (PDE). The decrease of intracellular cGMP concentrations causes the ion channels within the outer segment of the rod or cone to close, thus causing membrane hyperpolarization and, eventually, signal transmission. Rhodopsin and quote;s activity is believed to be shut off by its phosphorylation followed by binding of the soluble protein arrestin.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
1D4: a versatile epitope tag for the purification and characterization of expressed membrane and soluble proteins.
MA1-722 was used in immunocytochemistry and western blot to describe various immunochemical procedures used for the detection, purification, and localization of 1D4-tagged proteins
|Molday LL,Molday RS||Methods in molecular biology (Clifton, N.J.) (1177:1)||2014|
TSPAN12 regulates retinal vascular development by promoting Norrin- but not Wnt-induced FZD4/beta-catenin signaling.
MA1-722 was used in western blot to study the mechanism by which TSPAN12 regulates retinal vascular development
|Junge HJ,Yang S,Burton JB,Paes K,Shu X,French DM,Costa M,Rice DS,Ye W||Cell (139:299)||2009|
High levels of retinal membrane docosahexaenoic acid increase susceptibility to stress-induced degeneration.
MA1-722 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|
Delayed loss of cone and remaining rod photoreceptor cells due to impairment of choroidal circulation after acute light exposure in rats.
MA1-722 was used in western blot to investigate the effects of acute light exposure on rat retina and choroid
|Tanito M,Kaidzu S,Anderson RE||Investigative ophthalmology and visual science (48:1864)||2007|
Alleviation of constant-light-induced photoreceptor degeneration by adaptation of adult albino rat to bright cyclic light.
MA1-722 was used in western blot to demonstrate the rapid response of adult rat retina to light change.
|Li F,Cao W,Anderson RE||Investigative ophthalmology and visual science (44:4968)||2003|
RPGRIP1s with distinct neuronal localization and biochemical properties associate selectively with RanBP2 in amacrine neurons.
MA1-722 was used in western blot to investigate the distribution of RPGRIP1s and their association with RanBP2 in amacrine neurons
|Castagnet P,Mavlyutov T,Cai Y,Zhong F,Ferreira P||Human molecular genetics (12:1847)||2003|
Rpe65 is necessary for production of 11-cis-vitamin A in the retinal visual cycle.
MA1-722 was used in western blot to study the role of RPE65 in 11-cis-vitamin A production in retinal visual cycle
|Redmond TM,Yu S,Lee E,Bok D,Hamasaki D,Chen N,Goletz P,Ma JX,Crouch RK,Pfeifer K||Nature genetics (20:344)||1998|
Reproducible and sustained regulation of G¿s signalling using a metazoan opsin as an optogenetic tool.
MA1-722 was used in immunocytochemistry to evaluate a novel optogenetic tool for the study of G protein-coupled receptor signalling
|Bailes HJ,Zhuang LY,Lucas RJ||PloS one (7:null)||2012|
Disease sequence from mutant rhodopsin allele to rod and cone photoreceptor degeneration in man.
MA1-722 was used in immunocytochemistry to study the mechanism for rod and cone photoreceptor degeneration from rhodopsin mutations
|Cideciyan AV,Hood DC,Huang Y,Banin E,Li ZY,Stone EM,Milam AH,Jacobson SG||Proceedings of the National Academy of Sciences of the United States of America (95:7103)||1998|
A neural-specific F-box protein Fbs1 functions as a chaperone suppressing glycoprotein aggregation.
MA1-722 was used in immunoprecipitation to investigate the function of a neural-specific F-box protein Fbs1 in glycoprotein aggregation.
|Yoshida Y,Murakami A,Iwai K,Tanaka K||The Journal of biological chemistry (282:7137)||2007|
Antigen-antibody interaction. Synthetic peptides define linear antigenic determinants recognized by monoclonal antibodies directed to the cytoplasmic carboxyl terminus of rhodopsin.
MA1-722 was used in ELISA to investigate the specificity of monoclonal anti-rhodopsin antibodies
|Hodges RS,Heaton RJ,Parker JM,Molday L,Molday RS||The Journal of biological chemistry (263:11768)||1988|