Overview

  • Product name

    Anti-IL-10 antibody [EPR1114]
    See all IL-10 primary antibodies
  • Description

    Rabbit monoclonal [EPR1114] to IL-10
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WBmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human IL-10 aa 1-100. The exact sequence is proprietary.
    Database link: P22301
    (Peptide available as ab169704)

  • Positive control

    • Flow Cyt: HeLa cells; Human PBMCs. WB: recombinant IL-10 protein, Ramos, THP1 and THP1 treated with LPS cell lysates ICC/IF: Human peripheral blood mononuclear 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 ab133575 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 1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/50 - 1/250.
WB 1/1000 - 1/10000. Detects a band of approximately 18 kDa (predicted molecular weight: 20 kDa).Can be blocked with IL-10 peptide (ab169704).
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function

      Inhibits the synthesis of a number of cytokines, including IFN-gamma, IL-2, IL-3, TNF and GM-CSF produced by activated macrophages and by helper T-cells.
    • Tissue specificity

      Produced by a variety of cell lines, including T-cells, macrophages, mast cells and other cell types.
    • Sequence similarities

      Belongs to the IL-10 family.
    • Cellular localization

      Secreted.
    • Information by UniProt
    • Database links

    • Alternative names

      • CSIF antibody
      • Cytokine synthesis inhibitory factor antibody
      • GVHDS antibody
      • IL 10 antibody
      • IL-10 antibody
      • IL10 antibody
      • IL10_HUMAN antibody
      • IL10A antibody
      • Interleukin 10 antibody
      • Interleukin-10 antibody
      • MGC126450 antibody
      • MGC126451 antibody
      • T-cell growth inhibitory factor antibody
      • TGIF antibody
      see all

    Images

    • Overlay histogram showing HeLa cells fixed in 4% PFA and stained with purified ab133575 at a dilution of 1 in 300 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
    • Unpurified ab133575 staining IL-10 in human peripheral blood mononuclear cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Saponin in PBS and blocked with 4% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/100) was used as the secondary antibody.

      See Abreview

    • Lane 1 : Anti-IL-10 antibody [EPR1114] (ab133575) (purified)
      Lanes 2-3 : purified

      Lane 1 : THP-1 treated with LPS cell lysate
      Lane 2 : untreated THP-1 cell lysate
      Lane 3 : Ramos cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 20 kDa
      Observed band size: 18 kDa
      why is the actual band size different from the predicted?



      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • Immunofluorescence staining of Molt-4 cells with purified ab133575 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab133575 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

    • All lanes : Anti-IL-10 antibody [EPR1114] (ab133575) at 1/1000 dilution (unpurified)

      Lane 1 : LPS-treated THP1 cell lysate
      Lane 2 : THP1 cell lysate
      Lane 3 : Ramos cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 20 kDa
      Observed band size: 18 kDa why is the actual band size different from the predicted?

    • Anti-IL-10 antibody [EPR1114] (ab133575) at 1/1000 dilution (unpurified) + Recombinant IL-10 protein at 0.005 µg

      Secondary
      Standard HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 20 kDa
      Observed band size: 18 kDa why is the actual band size different from the predicted?

    • Flow cytometry analysis of permeabilized HeLa cells labelling IL-10 using unpurified ab133575 at a 1/100 dilution (red) or a control rabbit IgG antibody (green).

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Flow cytometric analysis of human PBMCs (peripheral blood mononuclear cells) treated with PHA labeling IL-10 with ab133575 at 1/100 dilution (green), followed by Goat anti-rabbit Alexa Fluor® 488.

      1 (dashed line) = isotype control, 2 (black line) = PBMCs untreated, 3 (green line) = PBMCs treated with PHA.

      See Abreview

    References

    This product has been referenced in:

    • Keuper M  et al. Activated macrophages control human adipocyte mitochondrial bioenergetics via secreted factors. Mol Metab 6:1226-1239 (2017). Read more (PubMed: 29031722) »
    • Raha AA  et al. Neuroprotective Effect of TREM-2 in Aging and Alzheimer's Disease Model. J Alzheimers Dis N/A:N/A (2016). IHC ; Mouse . Read more (PubMed: 27662313) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rat Tissue sections (Brain)
    Permeabilization
    Yes - 0.1 M PBS with 3% Triton X
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Paraformaldehyde

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Aug 20 2015

    Application
    Flow Cytometry
    Sample
    Human Cell (PBMCs (peripheral blood mononuclear cells))
    Permeabilization
    Yes - 0,1% Saponin /1% FCS/ PBS
    Gating Strategy
    no gate
    Specification
    PBMCs (peripheral blood mononuclear cells)
    Preparation
    Cell harvesting/tissue preparation method: isolated by gradient centrifugation
    Sample buffer: 0,1% Saponin /1% FCS/ PBS
    Fixation
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 27 2014

    Application
    Western blot
    Sample
    Human Recombinant protein (see below)
    Gel Running Conditions
    Reduced Non-Denaturing (Native) (5-16% gel)
    Loading amount
    10 µg
    Specification
    see below
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Oct 27 2014

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (PBMCs (peripheral blood mononuclear cells))
    Permeabilization
    Yes - 0,1% Saponin/PBS
    Specification
    PBMCs (peripheral blood mononuclear cells)
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 22 2014

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