Overview

  • 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.

  • Tested applications

    Suitable for: IHC-FoFr, IHC-Fr, ELISA, ICC, IP, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human, Zebrafish, Amphibian
    Predicted to work with: Rabbit
  • Immunogen

    Other Immunogen Type corresponding to Bovine Rhodopsin.

  • Epitope

    The epitope for this antibody has been localized to the C-terminal nine amino acids of bovine rhodopsin known as the 1D4 epitope.

Properties

  • 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...
  • 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

Applications

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
IHC-FoFr 1/1000. PubMed: 19587120
IHC-Fr Use at an assay dependent concentration. PubMed: 22743318
ELISA Use at an assay dependent concentration.
ICC 1/1000.
IP Use at an assay dependent concentration.

Use at an assay dependent dilution.

WB 1/1000. Detects a band of approximately 40 kDa.
IHC-P 1/100 - 1/1000.

Target

  • 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
    modifications

    Phosphorylated 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
  • Database links

  • Alternative names

    • CSNBAD1 antibody
    • MGC138309 antibody
    • MGC138311 antibody
    • OPN 2 antibody
    • OPN2 antibody
    • opsd antibody
    • OPSD_HUMAN antibody
    • opsin 2 antibody
    • Opsin 2 rod pigment antibody
    • Opsin-2 antibody
    • Opsin2 antibody
    • Retinitis Pigmentosa 4 antibody
    • Retinitis pigmentosa 4 autosomal dominant antibody
    • RHO antibody
    • Rhodopsin antibody
    • RP 4 antibody
    • RP4 antibody
    see all

Images

  • Deletion of Arf4 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.

  • 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.

  • 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.

  • Immunohistochemistry (Frozen sections) image of anti-Rhodopsin antibody (ab5417) staining on a Zebrafish eye.

    This tissue section was stained with anti-Rhodopsin antibody (green) and Red Opsin antibody (red) for 16 hours at 4°C. Nuclei were stained with DAPI (blue). This image suggests that ab5417 detects Red Opsin and not Rhodopsin in the Zebrafish. For more information please see Yin J. et al., 2012.

    See Abreview

  • 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.

  • 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?

References

This product has been referenced in:

  • Shrestha R  et al. Aberrant hiPSCs-Derived from Human Keratinocytes Differentiates into 3D Retinal Organoids that Acquire Mature Photoreceptors. Cells 8:N/A (2019). Read more (PubMed: 30634512) »
  • Cao X  et al. Factors governing the transduction efficiency of adeno-associated virus in the retinal ganglion cells following intravitreal injection. Gene Ther N/A:N/A (2019). Read more (PubMed: 30692605) »
See all 44 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Non-reduced Denaturing (12%)
Sample
Human Cell lysate - whole cell (neuroblast)
Specification
neuroblast
Blocking step
BSA as blocking agent for 50 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 17 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Sample
Zebrafish Tissue sections (Long double cone (red cone) photoreceptor cells)
Specification
Long double cone (red cone) photoreceptor cells
Permeabilization
Yes - PBST
Fixative
Paraformaldehyde

Dr. Jun Yin

Verified customer

Submitted Jul 02 2013

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