Recombinant
RabMAb

Recombinant Anti-RBP4 antibody [EP3657] (ab109193)

Overview

  • Product name

    Anti-RBP4 antibody [EP3657]
    See all RBP4 primary antibodies
  • Description

    Rabbit monoclonal [EP3657] to RBP4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human RBP4 aa 100-200. The exact sequence is proprietary.

  • Positive control

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

     

     

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109193 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
WB 1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function

      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.
    • Involvement in disease

      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.
    • Sequence similarities

      Belongs to the calycin superfamily. Lipocalin family.
    • Cellular localization

      Secreted.
    • Information by UniProt
    • Database links

    • Alternative names

      • 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

    Images

    • 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
    • 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
    • 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
    • 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 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.

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

    References

    This product has been referenced in:

    See all 5 Publications for this product

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    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

    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

    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.
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