Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸

Order now and get it on Friday March 05, 2021

Key features and details

Mouse monoclonal [1D4] to Rhodopsin
Suitable for: WB, IHC-P
Reacts with: Mouse, Human
Isotype: IgG1

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 & Species

Application Species
IHC-P
Mouse

Human

WB
Human

See all applications and species data
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.
Positive control
- WB: HL60 whole cell lysate IHC-P: Human and mouse retinal tissue
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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Application	Species
IHC-P	Mouse Human
WB	Human

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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rhodopsin antibody [1D4] (ab5417) Pearring et al PLoS Genet. 2017 Apr 14;13(4):e1006740. doi: 10.1371/journal.pgen.1006740. eCollection 2017 Apr. Fig 3. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

Deletion of Arf4 mice from the retina does not disrupt rhodopsin localization or photoreceptor morphology.

E. Arf4 immunostaining in Arf4^flox/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 Arf4^flox/CagCreER experimental and control retinal cross-sections. Eyes were collected at P34. Scale bar = 10 μm.

G. Comparative analysis of photoreceptor morphology in Arf4^flox/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.
Western blot - Anti-Rhodopsin antibody [1D4] (ab5417)

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?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rhodopsin antibody [1D4] (ab5417)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rhodopsin antibody [1D4] (ab5417)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rhodopsin antibody [1D4] (ab5417)

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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rhodopsin antibody [1D4] (ab5417)

Immunohistochemical analysis of paraffin-embedded humanretinal 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.

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