Overview

  • Product name

    Anti-Rhodopsin antibody [Rho 4D2]
    See all Rhodopsin primary antibodies
  • Description

    Mouse monoclonal [Rho 4D2] to Rhodopsin
  • Host species

    Mouse
  • Tested applications

    Suitable for: Flow Cyt, WB, IP, ELISA, IHC-P, IHC-Fr, ICCmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human, Pig, Xenopus laevis
    Does not react with: Drosophila melanogaster
  • Immunogen

    Recombinant fragment corresponding to Bovine Rhodopsin (N terminal).
    Database link: P02699

Properties

Applications

Our Abpromise guarantee covers the use of ab98887 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
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

WB 1/1000. Predicted molecular weight: 39 kDa.
IP Use at an assay dependent concentration. 1-2 µg per 100-500µg of protein.
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.

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

  • Immunohistochemistry analysis staining Rhodopsin in mouse retina with ab98887 at 1:1000. Secondary Antibody: FITC Goat Anti-Mouse (green). Counterstain: DAPI (blue) nuclear stain.

  • Overlay histogram showing Y79 cells stained with ab98887 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab98887, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Y79 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:

  • Daruich A  et al. Iron is neurotoxic in retinal detachment and transferrin confers neuroprotection. Sci Adv 5:eaau9940 (2019). Read more (PubMed: 30662950) »
  • Zou T  et al. Organoid-derived C-Kit+/SSEA4- human retinal progenitor cells promote a protective retinal microenvironment during transplantation in rodents. Nat Commun 10:1205 (2019). Read more (PubMed: 30872578) »
See all 12 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting Abcam and thank you for sending the details of your protocol as well as the image. These have been very helpful.

After reviewing your protocol, I would like to suggest a few tips which may help reduce the upper dark areas and increase the band quality for Rhodopsin.
First I would recommend changing your blocking procedures. It may be best for this product if your initial blocking and primary antibody incubation are performed using 3% BSA in TBST. This should be followed by use of a casein based commercial block for incubating your secondary antibody. We have seen that this may dramatically reduce background in western blot. We do carry this type of casein based blocking reagent, ab64234 (https://www.abcam.com/Background-Reducing-Buffer-ab64234-support.html).

Second, you may wish to reduce the concentration of secondary antibody that you use. This product has a large range 1:2000-1:20,000 against which it has been used effectively.

Finally I would suggest increasing the concentration of detergent in your TBST, if you currently use 0.1% Tween20 use 0.25% Tween20 instead.

Read More
Application
Western blot
Sample
Bird Tissue lysate - whole (Pigeon Retina, no boiling of sample prior to loadi)
Gel Running Conditions
Non-reduced Denaturing (7.5% Tris-Glycine)
Loading amount
20 µg
Specification
Pigeon Retina, no boiling of sample prior to loadi
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Tobias Hochstöger

Verified customer

Submitted Sep 15 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Retina)
Gel Running Conditions
Reduced Denaturing (12% Gel)
Loading amount
2 µg
Specification
Retina
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Onkar Sawant

Verified customer

Submitted Mar 04 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Eye)
Antigen retrieval step
None
Permeabilization
Yes - Tween 0.1%
Specification
Eye
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2 · Temperature: 24°C
Fixative
Paraformaldehyde

Onkar Sawant

Verified customer

Submitted Jan 28 2016

Answer

Thank you for contacting us.

The band size seems to be correct. This product has a predicted weight of 39kDa although you band seems a bit smaller this is common when using in real world settings as compared to predictions. Post translational modifications are a common cause of this as well as the inherent inaccuracies of protein marker standards. These are often labeled as 'apparent' size because they may be off by 3-30kDa.
In regards to further data and publications, to our knowledge this product has not been published with. We always include any information that we may have regarding publications, reviews or any information that we about the antibody on our website.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this recommended product was not correct for your assay and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number I have also attached a pdf file with our recommended protocol for stripping western blots. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

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