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Anti-Rhodopsin 抗体 [Rho 4D2] (ab98887)

Reviews (3)Q&A (3)References (29)
Immunohistochemistry (Frozen sections) - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)
  • Western blot - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)
  • Western blot - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)
  • Flow Cytometry - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)

Key features and details

  • Mouse monoclonal [Rho 4D2] to Rhodopsin
  • Suitable for: Flow Cyt, ELISA, ICC, IHC-P, IHC-Fr, IP, WB
  • Reacts with: Mouse, Rat, Cow, Human, Pig, Xenopus laevis
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-Rhodopsin antibody [Rho 4D2]
    Rhodopsin 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [Rho 4D2] to Rhodopsin

  • 由来種

    Mouse

  • アプリケーション

    適用あり: Flow Cyt, ELISA, ICC, IHC-P, IHC-Fr, IP, WBmore details

  • 種交差性

    交差種: Mouse, Rat, Cow, Human, Pig, Xenopus laevis
    非交差種: Drosophila melanogaster

  • 免疫原

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

  • ポジティブ･コントロール

    • Flow Cyt: Y79 cells. IHC-Fr: Mouse retina tissue. WB: A549 cell lysate. Bovine photoreceptor membrane tissue lysate.

  • 特記事項

    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

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab98887の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ELISA
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.

1-2 ug per 100-500ug of protein.
WB (2)
1/1000. Predicted molecular weight: 38 kDa.
特記事項
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ELISA
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.

1-2 ug per 100-500ug of protein.
WB
1/1000. Predicted molecular weight: 38 kDa.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    Rod shaped photoreceptor cells which mediates vision in dim light.

  • 関連疾患

    Retinitis pigmentosa 4
    Night blindness, congenital stationary, autosomal dominant 1

  • 配列類似性

    Belongs to the G-protein coupled receptor 1 family. Opsin subfamily.

  • 翻訳後修飾

    Phosphorylated on some or all of the serine and threonine residues present in the C-terminal region.
    Contains one covalently linked retinal chromophore.

  • 細胞内局在

    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

  • 参照データベース

    see all

  • 別名

    • 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

画像

  • Immunohistochemistry (Frozen sections) - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)
    Immunohistochemistry (Frozen sections) - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)

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

    OS - photoreceptor outer segment; IS – inner segment; ONL – outer nuclear layer; OPL – outer plexiform layer; INL – inner nuclear layer; IPL – inner plexiform layer; GCL – ganglion cell layer.

  • Western blot - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)
    Western blot - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)
    All lanes : Anti-Rhodopsin antibody [Rho 4D2] (ab98887) at 1/1000 dilution

    Lane 1 : Bovine photoreceptor membrane tissue lysate at 10 µg
    Lane 2 : Bovine photoreceptor membrane tissue lysate at 5 µg
    Lane 3 : Bovine photoreceptor membrane tissue lysate at 2.5 µg

    Predicted band size: 38 kDa
    Observed band size: 110,35,75 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)
    Western blot - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)
    Anti-Rhodopsin antibody [Rho 4D2] (ab98887) at 1/1000 dilution (2.5 hours at room temperature with shaking) + A549 (human lung carcinoma cell line) whole cell lysate at 15 µg

    Secondary
    Goat Anti-Mouse IgG (HRP) for 1 hour at room temperature with shaking at 1/1000 dilution

    Predicted band size: 38 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?



    Band appears at ~75 kDa indicating detection of the Rhodopsin dimer.

     

    Block: 5% Skim Milk Powder in TBST.

     

    Color Development: Chemiluminescent for HRP for 5 min at room temperature.

  • Flow Cytometry - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)
    Flow Cytometry - Anti-Rhodopsin antibody [Rho 4D2] (ab98887)

    Overlay histogram showing Y79 (Human retinoblastoma cell line) 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.

参考文献 (29)

ab98887 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab98887 は 29 報の論文で使用されています。

  • Sheng W  et al. Immunohistological Localization of Mel1a Melatonin Receptor in Pigeon Retina. Nat Sci Sleep 13:113-121 (2021). PubMed: 33574722
  • Dahl TM  et al. Effect of conditional deletion of cytoplasmic dynein heavy chain DYNC1H1 on postnatal photoreceptors. PLoS One 16:e0248354 (2021). PubMed: 33705456
  • Almarhoun M  et al. Overexpression of STARD3 attenuates oxidized LDL-induced oxidative stress and inflammation in retinal pigment epithelial cells. Biochim Biophys Acta Mol Cell Biol Lipids 1866:158927 (2021). PubMed: 33771709
  • Chen YW  et al. The Functional Vision Protection Effect of Danshensu via Dopamine D1 Receptors: In Vivo Study. Nutrients 13:N/A (2021). PubMed: 33803057
  • Crouzier L  et al. Loss of Pde6a Induces Rod Outer Segment Shrinkage and Visual Alterations in pde6aQ70X Mutant Zebrafish, a Relevant Model of Retinal Dystrophy. Front Cell Dev Biol 9:675517 (2021). PubMed: 34095146
View all Publications for this product

レビューと Q&A

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1-6 of 6 Abreviews or Q&A

Western blot abreview for Anti-Rhodopsin antibody [Rho 4D2]

 Excellent
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Tobias Hochstöger

Verified customer

投稿 Sep 15 2017

Western blot abreview for Anti-Rhodopsin antibody [Rho 4D2]

 Excellent
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Onkar Sawant

Verified customer

投稿 Mar 04 2016

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Rhodopsin antibody [Rho 4D2]

 Excellent
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Onkar Sawant

Verified customer

投稿 Jan 28 2016

Question

I need help again, I send you the file of my WB with the Rhodopsin ab98887 you report the ab is around 39kDa if you see the image of the wb the band maybe I think it is less than 37kDa, what do you think? And I am very surprise how the ab are up the 50 to 250kDa? Is not very clean ab? What are your suggestions? This is what I did: SDS 10%, transfer to Nitrocellulose, Blocking for 1hr at RT with 5%Milk on TBSt, the primary ab Rhodopsin ab98887 at 1:1000 on 1% Milk overnight, 4 washes with TBSTt every 5 min, incubate with secondary ab98693 at 1:2000 on 1% Milk for 1hr, 6washes of 5min with TBST, and I did ECL the exposition time was very fast like 10 sec, more than that is get it so dark.

Read More
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

Question

Hi, I'll try again all this changes to see if I can improve the quality of the band of Rhodopsin, but what about the size of the band? Is the correct size? And Can you please send me if you have reviews using this ab, Do you have publications with this ab? Thanks,

Read More
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.

Read More

Question

Phoned because they have seen no bands in WB with these products.

Read More
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.

Read More

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