Anti-Rhodopsin antibody [1D4] (ab5417)
Key features and details
- Mouse monoclonal [1D4] to Rhodopsin
- Suitable for: ELISA, IHC-FoFr, IP, WB, IHC-Fr, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Cow, Human, Zebrafish, Amphibian
- Isotype: IgG1
Overview
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Product name
Anti-Rhodopsin antibody [1D4]
See all Rhodopsin primary antibodies -
Description
Mouse monoclonal [1D4] to Rhodopsin -
Host species
Mouse -
Specificity
ab5417 detects Rhodopsin from human and bovine retinal samples. Data from Yin J et al., 2012 (PMID 22743318) indicates that in Zebrafish ab5417 appears to recognize Red Opsin rather than Rhodopsin.
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Tested applications
Suitable for: ELISA, IHC-FoFr, IP, WB, IHC-Fr, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Cow, Human, Zebrafish, Amphibian
Predicted to work with: Rabbit -
Immunogen
Tissue, cells or virus corresponding to Bovine Rhodopsin. Bleached bovine ROS [rod outer segment] disk membranes
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Epitope
The epitope for this antibody has been localized to the C-terminal nine amino acids of bovine rhodopsin known as the 1D4 epitope. -
Positive control
- WB: HL60 whole cell lysate IHC-P: Human and mouse retinal tissue
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituents: 99% PBS, 0.1% BSA -
Concentration information loading...
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Purity
Protein G purified -
Primary antibody notes
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’s activity is believed to be shut off by its phosphorylation followed by binding of the soluble protein arrestin. -
Clonality
Monoclonal -
Clone number
1D4 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab5417 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ELISA |
Use at an assay dependent concentration.
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IHC-FoFr |
Use at an assay dependent concentration. PubMed: 19587120
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IP |
Use at an assay dependent concentration.
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WB | (2) |
1/100 - 1/1000. Detects a band of approximately 40 kDa.
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IHC-Fr | (1) |
Use at an assay dependent concentration. PubMed: 22743318
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IHC-P |
1/100 - 1/1000.
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ICC/IF | (1) |
Use at an assay dependent concentration.
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Notes |
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ELISA
Use at an assay dependent concentration. |
IHC-FoFr
Use at an assay dependent concentration. PubMed: 19587120 |
IP
Use at an assay dependent concentration. |
WB
1/100 - 1/1000. Detects a band of approximately 40 kDa. |
IHC-Fr
Use at an assay dependent concentration. PubMed: 22743318 |
IHC-P
1/100 - 1/1000. |
ICC/IF
Use at an assay dependent concentration. |
Target
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Function
Photoreceptor required for image-forming vision at low light intensity. Required for photoreceptor cell viability after birth. Light-induced isomerization of 11-cis to all-trans retinal triggers a conformational change leading to G-protein activation and release of all-trans retinal. -
Tissue specificity
Rod shaped photoreceptor cells which mediates vision in dim light. -
Involvement in disease
Retinitis pigmentosa 4
Night blindness, congenital stationary, autosomal dominant 1 -
Sequence similarities
Belongs to the G-protein coupled receptor 1 family. Opsin subfamily. -
Post-translational
modificationsPhosphorylated on some or all of the serine and threonine residues present in the C-terminal region.
Contains one covalently linked retinal chromophore. -
Cellular localization
Membrane. Synthesized in the inner segment (IS) of rod photoreceptor cells before vectorial transport to the rod outer segment (OS) photosensory cilia. - Information by UniProt
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Database links
- Entrez Gene: 509933 Cow
- Entrez Gene: 100307115 Human
- Entrez Gene: 6010 Human
- Entrez Gene: 212541 Mouse
- Entrez Gene: 100009200 Rabbit
- Entrez Gene: 24717 Rat
- Entrez Gene: 30295 Zebrafish
- Omim: 180380 Human
see all -
Alternative names
- CSNBAD1 antibody
- MGC138309 antibody
- MGC138311 antibody
see all
Images
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Deletion of Arf4 mice from the retina does not disrupt rhodopsin localization or photoreceptor morphology.
E. Arf4 immunostaining in Arf4flox/CagCreER experimental and control retinal cross-sections. Image of the photoreceptor IS where the biosynthetic membranes are localized. Eyes were collected at P34. Scale bar = 10 μm.
F. Rhodopsin immunostaining in Arf4flox/CagCreER experimental and control retinal cross-sections. Eyes were collected at P34. Scale bar = 10 μm.
G. Comparative analysis of photoreceptor morphology in Arf4flox/CagCreER experimental and control retinal cross-sections. Eyes were collected at P41. Scale bar = 20 μm.
OS = outer segment, IS = inner segment, ONL = outer nuclear layer, OPL = outer plexiform layer.
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Anti-Rhodopsin antibody [1D4] (ab5417) at 1/500 dilution + HL60 (Human promyelocytic leukemia cell line) cell lysate at 25 µg
Observed band size: 40 kDa why is the actual band size different from the predicted? -
Immunohistochemical analysis of formalin-fixed mouse retinal tissue, labeling rhodopsin with ab5417 at a 1:50 dilution in 3% BSA-PBS solution and incubated at 4°C overnight in a high humidity environment.
A DyLight® 488 secondary antibody was used (green) incubated at room temperature in the dark. The tissue was counterstained with DAPI against DNA, showing nuclear compartments. Prior to staining the formalin-fixed tissue was permeabilized with 0.1% Triton X-100 in TBS for between 5 and 10 minutes, then blocked with 3% BSA-PBS for 30 minutes at room temperature.
The left image is a negative control with only the secondary antibody and the right image is in the presence of ab5417 and the secondary.
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Immunohistochemical analysis of formalin-fixed human retinal tissue, labeling rhodopsin with ab5417 at a 1:50 dilution in 3% BSA-PBS solution and incubated at 4°C overnight in a high humidity environment.
A DyLight® 488 secondary antibody was used (green) incubated at room temperature in the dark. The tissue was counterstained with DAPI against DNA, showing nuclear compartments. Prior to staining the formalin-fixed tissue was permeabilized with 0.1% Triton X-100 in TBS for between 5 and 10 minutes, then blocked with 3% BSA-PBS for 30 minutes at room temperature.
The left image is a negative control with only the secondary antibody and the right image is in the presence of ab5417 and the secondary.
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Immunohistochemical analysis of paraffin-embedded mouse retinal tissue labeling Rhodopsin with ab5417.
Secondary used was HRP conjugated. Prior preparation was initiated by antigen retrival using 10mM sodium citrate at pH 6.0, then the sample was microwaved for 8 to 15 minutes. Subsequent to retrival the retinal tissue was blocked for 15 minutes at room temperature with 3% hydrogen peroxide. The sample was then incubated with ab5417 in 3% BSA-PBS at 4°C at a dilution of 1:1000, overnight. Hematoxylin was used to counterstain the tissue.
The left side of the image is shown as a negative control and is the tissue in the absence of ab5417, the right side is in the prescence of the counterstain, ab5417 and the HRP conjugated secondary.
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Immunohistochemical analysis of paraffin-embedded human retinal tissue labeling Rhodopsin with ab5417.
Secondary used was HRP conjugated. Prior preparation was initiated by antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Subsequent to retrival the retinal tissue was blocked in 3% H2O2-methanol for 15 min at room temperature. The sample was then incubated with ab5417 in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C, overnight. Hematoxylin was used to counterstain the tissue.
The left side of the image is shown as a negative control and is the tissue in the absence of ab5417, the right side is in the prescence of the counterstain, ab5417 and the HRP conjugated secondary.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (94)
ab5417 has been referenced in 94 publications.
- Li X et al. Targeting long noncoding RNA-AQP4-AS1 for the treatment of retinal neurovascular dysfunction in diabetes mellitus. EBioMedicine 77:103857 (2022). PubMed: 35172268
- Zhang YF et al. Frequency cumulative effect of subthreshold energy laser-activated remote phosphors irradiation on visual function in guinea pigs. Int J Ophthalmol 15:213-220 (2022). PubMed: 35186679
- McDowell RJ et al. Divergent G-protein selectivity across melanopsins from mice and humans. J Cell Sci 135:N/A (2022). PubMed: 35274137
- Mathew D et al. Circadian rhythm disruption results in visual dysfunction. FASEB Bioadv 4:364-378 (2022). PubMed: 35664832
- Vagni P et al. POLYRETINA restores light responses in vivo in blind Göttingen minipigs. Nat Commun 13:3678 (2022). PubMed: 35760775