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リコンビナントRabMAb

Anti-RBP4 抗体 [EP3657] (ab109193)

Submit a review Q&A (3)References (10)
Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
  • Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
  • Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
  • Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
  • Immunocytochemistry/ Immunofluorescence - Anti-RBP4 antibody [EP3657] (ab109193)
  • Immunocytochemistry/ Immunofluorescence - Anti-RBP4 antibody [EP3657] (ab109193)
  • Anti-RBP4 antibody [EP3657] (ab109193)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP3657] to RBP4
  • Suitable for: WB, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-RBP4 antibody [EP3657]
    RBP4 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EP3657] to RBP4

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, ICC/IFmore details
    適用なし: IHC-P

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: Human plasma, HepG2 cell lysates, Human fetal kidney, Mouse serum and Rat serum. ICC/IF: HeLa cells

  • 特記事項

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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ab109193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
WB
1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
ICC/IF
1/100 - 1/250.
特記事項
WB
1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
ICC/IF
1/100 - 1/250.
追加情報
Is unsuitable for IHC-P.

ターゲット情報

  • 機能

    Delivers retinol from the liver stores to the peripheral tissues. In plasma, the RBP-retinol complex interacts with transthyretin, this prevents its loss by filtration through the kidney glomeruli.

  • 関連疾患

    Defects in RBP4 are a cause of retinol-binding protein deficiency (RBP deficiency) [MIM:180250]. This condition causes night vision problems. It produces a typical 'fundus xerophthalmicus', featuring a progressed atrophy of the retinal pigment epithelium.

  • 配列類似性

    Belongs to the calycin superfamily. Lipocalin family.

  • 細胞内局在

    Secreted.
  • Information by UniProt

  • 参照データベース

    • 別名

      • OTTHUMP00000020114 antibody
      • OTTHUMP00000020115 antibody
      • OTTHUMP00000020116 antibody
      • Plasma retinol binding protein 4 antibody
      • Plasma retinol-binding protein antibody
      • Plasma retinol-binding protein(1-176) antibody
      • PRBP antibody
      • PRO2222 antibody
      • RBP antibody
      • RBP4 antibody
      • RDCCAS antibody
      • RET4_HUMAN antibody
      • Retinol binding protein 4 antibody
      • Retinol binding protein 4 interstitial antibody
      • Retinol binding protein 4 plasma antibody
      see all

    画像

    • Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
      Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
      All lanes : Anti-RBP4 antibody [EP3657] (ab109193) at 1/5000 dilution

      Lane 1 : HepG2 (human hepatocellular carcinoma) whole cell lysate
      Lane 2 : Human fetal kidney

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 23 kDa
      Observed band size: 23 kDa



      Blocking and diluting buffer 5% NFDM/TBST
    • Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
      Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
      Anti-RBP4 antibody [EP3657] (ab109193) at 1/5000 dilution + Mouse serum at 20 µg

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 23 kDa
      Observed band size: 23 kDa



      Blocking and diluting buffer 5% NFDM/TBST
    • Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
      Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
      Anti-RBP4 antibody [EP3657] (ab109193) at 1/5000 dilution + Rat serum at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 23 kDa
      Observed band size: 23 kDa



      Blocking and diluting buffer 5% NFDM/TBST
    • Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
      Western blot - Anti-RBP4 antibody [EP3657] (ab109193)
      All lanes : Anti-RBP4 antibody [EP3657] (ab109193) at 1/1000 dilution (un-purified)

      Lane 1 : Human plasma lysate
      Lane 2 : HepG2 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 23 kDa
      Observed band size: 23 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-RBP4 antibody [EP3657] (ab109193)
      Immunocytochemistry/ Immunofluorescence - Anti-RBP4 antibody [EP3657] (ab109193)

      Immunocytochemistry/immunofluorescence staining of 100% methanol fixed; 0.1% triton X 100 permeabilized HeLa (human cervix adenocarcinoma) cells with purified ab109193 at dilution of 1/100. The secondary antibody used was Alexa Fluor® 488; goat anti-rabbit IgG (ab150077) at a dilution of 1/1000. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/1000) was used to stain tubulin along with ab150120 (AlexaFluor®594 goat anti-mouse secondary, 1/1000) shown in the top right hand panel. The negative controls are shown in the bottom middle and right hand panels- for negative control 1 rabbit primary antibody and anti-mouse secondary antibody (ab150120) was used. For negative control 2 mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) was used.

    • Immunocytochemistry/ Immunofluorescence - Anti-RBP4 antibody [EP3657] (ab109193)
      Immunocytochemistry/ Immunofluorescence - Anti-RBP4 antibody [EP3657] (ab109193)

      Immunofluorescent analysis of HeLa cells labelling RBP4 with un-purified ab109193 at a dilution of 1/100.

    • Anti-RBP4 antibody [EP3657] (ab109193)
      Anti-RBP4 antibody [EP3657] (ab109193)

    参考文献 (10)

    ab109193 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。

    ab109193 は 10 報の論文で使用されています。

    • Saeed A  et al. Impaired Hepatic Vitamin A Metabolism in NAFLD Mice Leading to Vitamin A Accumulation in Hepatocytes. Cell Mol Gastroenterol Hepatol 11:309-325.e3 (2021). PubMed: 32698042
    • Yoshida Y  et al. Alteration of circadian machinery in monocytes underlies chronic kidney disease-associated cardiac inflammation and fibrosis. Nat Commun 12:2783 (2021). PubMed: 33986294
    • Welles JE  et al. Retinol-binding protein 4 mRNA translation in hepatocytes is enhanced by activation of mTORC1. Am J Physiol Endocrinol Metab 320:E306-E315 (2021). PubMed: 33284085
    • Paul A  et al. Picomolar detection of retinol binding protein 4 for early management of type II diabetes. Biosens Bioelectron 128:122-128 (2019). PubMed: 30641454
    • Hashimoto N  et al. Involvement of the hepatic branch of the vagus nerve in the regulation of plasma adipokine levels in rats fed a high-fructose diet. J Nutr Biochem 71:90-97 (2019). PubMed: 31306952
    • Schuh CD  et al. Combined Structural and Functional Imaging of the Kidney Reveals Major Axial Differences in Proximal Tubule Endocytosis. J Am Soc Nephrol 29:2696-2712 (2018). PubMed: 30301861
    • Wang Y  et al. Adipokine RBP4 drives ovarian cancer cell migration. J Ovarian Res 11:29 (2018). PubMed: 29642915
    • Wang J  et al. Transcriptome Profiling Reveals PHLDA1 as a Novel Molecular Marker for Ischemic Cardiomyopathy. J Mol Neurosci N/A:N/A (2018). PubMed: 29736818
    • Dong S  et al. Involvement of RBP4 in all-trans retinoic acid induced cleft palate. Mol Med Rep 16:5915-5923 (2017). PubMed: 28849085
    • Kumar S  et al. Smoking accelerates pancreatic cancer progression by promoting differentiation of MDSCs and inducing HB-EGF expression in macrophages. Oncogene N/A:N/A (2014). WB ; Mouse . PubMed: 24909166

    レビューと Q&A

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    レビューを送る 質問を送る

    1-3 of 3 Abreviews or Q&A

    Question

    Regarding you questions, samples used were reduced and denatured in SDS sample buffer + B-ME and heated at 98 degrees for 5' prior loading. Lysis buffer was supplemented with protease inhibitors (mamalian protease inhibitor cocktail, Sigma). Rbp4 has been previously assessed by WB in mouse brain samples using non-abcam antibodies (see attached paper). If it is strictly necessary for obtaining another Rbp4 antibody for wb that crossreacts with mouse, or a refund, I can run another WB with mouse serum and kidney lysates along with my samples.

    Read More
    Answer


    I am happy to hear you have already reduced, denatured, and protected your samples from proteases. The only other attempt that I think is worth your time is running a WB with mouse serum and kidney lysates, especially since the wildtype levels of Rbp4 (when 30ug of whole brain cell lysates were loaded) barely produced a detectable WB band in the paper you referenced. If the antibody detects Rbp4 in the positive controls, then you know the antibody is properly detecting Rbp4 and that the levels in your sample may be very low, necessitating a much higher sample loading amount.

    I look forward to your reply and hearing whether or not the positive controls yield the expected Rbp4 band.

    Read More

    Question

    Inquiry: I recently purchased your anti-RBP4 antibody (ab109193) and used it for western blot analysis of mouse brain samples lysed in supplemented RIPA buffer. 20ug/lane were loaded in a 15-well acryl/bis gel and transferred to a nitrocellulose membrane at 100v for 90 min. Membrane was blocken in 5% milk in TBS-T prior to addition of primary antibody in blocking buffer at 1:1000 dilution (suggested dilution 1:1000-1:10,000). Membrane was incubated O/N at 4º with gentle agitation. Anti-rabbit secondary antibody (Jackson immunoresearch) was diluted at 1:10,000 in blocking buffer and added to the membrane, and incubated for 1h at RT. Detection was performed using ECL West Pico and West Dura (Pierce). No band was detected at the expected size (23kDa). Other bands (between 75 and 37kDa) were detected. films were exposed up to one hour. Rbp4 is expressed in the mouse brain according to the Allen institute atlas (http://mouse.brain-map.org/gene/show/19425). I would like a refund or a replacement for a different anti-RBP4 that crossreacts with mouse.Thank you very much.

    Read More
    Answer

    Thank you for contacting Abcam and providing your protocol information.

    I am sorry to hear that you have been experiencing issues when using this antibody. This product is covered by our Abpromise guarantee. According to Abpromise guarantee, if this product has been purchased within the last six months, we are happy to provide scientific support and to replace or refund the product should we not be able to help.

    Upon reviewing your protocol, I have a few additional questions/troubleshooting tips:
    1. Did you reduce and denature your samples? We recommend adding Beta-mercaptoethanol or DTT along with SDS and boiling with your sample 95-100oC for 5 minutes.
    2. Did you add protease and phosphatase inhibitors to your lysis buffer?
    3. Did you run a positive control? If you have access to human plasma, HeLa or HepG2 cells, or liver or kidney tissue these have all been used as positive controls for RBP4. While the mouse atlas may site specific examples of RBP4 expression in mouse brain, the actual RBP4 protein may be present at very low levels or not translated into actual protein. Gene expression does not guarantee translation into the endogenous RBP4 protein. A positive control will help determine if the antibody is properly detecting RBP4 or if the protein is not present in detectable levels in your sample.

    These will not only help me to fully understand the problem but are essential should further testing of this product be needed as well. Could you also provide the Abcam order confirmation number or the PO and date of purchase for this product?

    Read More

    Question

    Dear Sir/Madam,

    I have some questions about the RBP4 antibody whose catalog number isab109193. I'd like to know whether this antibody could be used as nutralizing antibody, and if so whether there is a protocal . Do you have anyRBP4 nutralizingantibody ?

    Thank you for your attention and expect for your prompt reply.

    Best Regards

    Read More
    Answer

    Thank you for contacting us.
    ab109193 has not been tested for neutralisation and we have no empirical data to support the application at the moment.
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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