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  1. Link

    il-10-antibody-jes5-2a5-ab33471.pdf

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Immunology Innate Immunity Cytokines Interleukins
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Anti-IL-10 antibody [JES5-2A5] (ab33471)

  • Datasheet
  • SDS
Reviews (6)Q&A (7)References (32)

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Key features and details

  • Rat monoclonal [JES5-2A5] to IL-10
  • Suitable for: ELISA, IHC-Fr, WB
  • Reacts with: Mouse, Rat
  • Isotype: IgG1

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Overview

  • Product name

    Anti-IL-10 antibody [JES5-2A5]
    See all IL-10 primary antibodies
  • Description

    Rat monoclonal [JES5-2A5] to IL-10
  • Host species

    Rat
  • Tested applications

    Suitable for: ELISA, IHC-Fr, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant full length protein corresponding to Mouse IL-10.

  • Positive control

    • WB: LPS treated mouse liver lysate. LPS treated mouse hepatocyte lysate. IHC-Fr: Rat brain tissue.
  • General notes

    ab33471 recognises mouse Interleukin 10 (IL-10). IL-10 is expressed by Ly 1 (B 1) B cells that express CD5 (Ly 1) and a subset of lectin stimulated T cells. Clone JES 5-2A5 is reported to neutralise the activity of murine IL-10.

    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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Storage buffer

    pH: 7.20
    Preservative: 0.09% Sodium azide
    Constituents: 0.87% Sodium chloride, 0.14% Monobasic dihydrogen sodium phosphate
  • Concentration information loading...
  • Purity

    Affinity purified
  • Purification notes

    Purified from TCS.
  • Primary antibody notes

    ab33471 recognises mouse Interleukin 10 (IL 10). IL 10 is expressed by Ly 1 (B 1) B cells that express CD5 (Ly 1) and a subset of lectin stimulated T cells. Clone JES 5-2A5 is reported to neutralise the activity of murine IL 10.
  • Clonality

    Monoclonal
  • Clone number

    JES5-2A5
  • Myeloma

    P3x63-Ag8.653
  • Isotype

    IgG1
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-IL-10 antibody [JES5-2A5] (ab189392)
  • Compatible Secondaries

    • Goat Anti-Rat IgG H&L (Alexa Fluor® 488) (ab150157)
    • Goat Anti-Rat IgG H&L (HRP) (ab205720)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rat IgG1, kappa monoclonal [RTK2071] - Isotype control (ab18412)
  • Recombinant Protein

    • Recombinant human IL-10 protein (Active) (ab259402)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab33471 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
ELISA
Use at an assay dependent concentration.
IHC-Fr (1)
Use at an assay dependent concentration.
WB (4)
Use at an assay dependent concentration. Predicted molecular weight: 20 kDa.

We recommend incubating the membrane overnight in primary antibody.

Notes
ELISA
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 20 kDa.

We recommend incubating the membrane overnight in primary antibody.

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.
  • Target information above from: UniProt accession P22301 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 16153 Mouse
    • Entrez Gene: 25325 Rat
    • SwissProt: P18893 Mouse
    • SwissProt: P29456 Rat
    • Unigene: 874 Mouse
    • Unigene: 9868 Rat
    • 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

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (32)

    Publishing research using ab33471? Please let us know so that we can cite the reference in this datasheet.

    ab33471 has been referenced in 32 publications.

    • Li N  et al. Dental pulp stem cells overexpressing hepatocyte growth factor facilitate the repair of DSS-induced ulcerative colitis. Stem Cell Res Ther 12:30 (2021). PubMed: 33413675
    • Liu H  et al. Adipose-derived mesenchymal stromal cell-derived exosomes promote tendon healing by activating both SMAD1/5/9 and SMAD2/3. Stem Cell Res Ther 12:338 (2021). PubMed: 34112236
    • Luo Y  et al. Inhibition of EZH2 (Enhancer of Zeste Homolog 2) Attenuates Neuroinflammation via H3k27me3/SOCS3/TRAF6/NF-?B (Trimethylation of Histone 3 Lysine 27/Suppressor of Cytokine Signaling 3/Tumor Necrosis Factor Receptor Family 6/Nuclear Factor-?B) in a Rat Model of Subarachnoid Hemorrhage. Stroke 51:3320-3331 (2020). PubMed: 32933418
    • Chen JJ  et al. Ribonuclease T2 from Aspergillus fumigatus promotes T helper type 2 responses through M2 polarization of macrophages. Int J Mol Med 46:718-728 (2020). PubMed: 32468025
    • Dumbuya JS  et al. G-CSF attenuates neuroinflammation and neuronal apoptosis via the mTOR/p70SK6 signaling pathway in neonatal Hypoxia-Ischemia rat model. Brain Res 1739:146817 (2020). PubMed: 32246916
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 13 Abreviews or Q&A

    Western blot abreview for Anti-IL-10 antibody [JES5-2A5]

    Poor
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Brain)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (12% gel)
    Loading amount
    40 µg
    Specification
    Brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. WeiHua Wu

    Verified customer

    Submitted Mar 25 2019

    Western blot abreview for Anti-IL-10 antibody [JES5-2A5]

    Poor
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain lysis)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Loading amount
    40 µg
    Specification
    Brain lysis
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Dr. WeiHua Wu

    Verified customer

    Submitted Mar 25 2019

    Western blot abreview for Anti-IL-10 antibody [JES5-2A5]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (mouse liver, mouse hepatocytes)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    25 µg
    Treatment
    LPS 5mg/kg iv for liver, 100 ng/ml for hepatocytes
    Specification
    mouse liver, mouse hepatocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 23 2017

    Immunohistochemistry (Frozen sections) abreview for Anti-IL-10 antibody [JES5-2A5]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    DR. Ruma Raha-Chowdhury

    Verified customer

    Submitted Jun 25 2015

    Western blot abreview for Anti-IL-10 antibody [JES5-2A5]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (liver)
    Gel Running Conditions
    Reduced Non-Denaturing (Native)
    Loading amount
    20 µg
    Specification
    liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 13 2014

    Question

    Customer kindly contacted us to find an anti-IL10 product to use in IHC-P/F in mouse. This product cannot react to human IL-10.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 12 2012

    Answer

    Thank you for contacting us.

    I have found some information regarding the anti-IL-10 antibodies which we spoke of. To begin, I have found that Mouse IL-10 and Human IL-10 have only a 73% homology which is especially poor in the region around aa13-aa70 of mouse IL-10 (http://www.uniprot.org/uniprot/P18893).

    The peptide from which ab9969 was created has 78% identity. Generally we do not expect an antibody to recognize a target if the homology is below 85% but this product is polyclonal and as such may be able to recognize human IL-10 in some lots.



    The immunogen used to generate ab33471 is E coli-expressed recombinant mouse IL-10. We have not mapped the epitope of this antibody so we do not know exactly where the antibody will bind. .However this is a monoclonal product and as stated above the homology between IL-1o mouse and human is below the threshold for antibody recognition. As such this may be an appropriate choice but we have not tested in-house for cross reactivity of this antibody andcannot guarantee that it will not recognize human IL-10.



    The same statements above can be repeated for ab25073. However as this is a mouse monoclonal and you wish to work in mouse tissue we would highly recommend use of theMouse on Mouse Polymer IHC Kit ab127055.



    AB33471 and ab9969 would also qualify for enrollment in our AbTrial testing program. For UNTESTED applications, we have established a testing discount program. Here is a brief description of how it works:

    The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount equal to the value of the product tested. The terms and conditions of this offer can be found here:https://www.abcam.com/AbTrial

    I have included links to each of these products below:

    AB9969:https://www.abcam.com/il10-antibody-ab9969.html

    AB33471:https://www.abcam.com/il10-antibody-jes5-2a5-ab33471.html

    AB25073:https://www.abcam.com/il10-antibody-2g101h7-ab25073.html

    Mouse on Mouse Kit:https://www.abcam.com/Mouse-on-Mouse-MOM-Polymer-IHC-Kit-ab127055.html

    I hope this information is helpful to you. If you are interested in testing one of these products in IHC-P please let me know. 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

    Read More

    Abcam Scientific Support

    Answered on Oct 13 2012

    Question


    I haveone questions on the Anti-IL10 antibody [JES5-2A5] (ab33471),could please help me:

    (1) which kind of tissue(from rat or mouse) could be used as positive control tissue when I do FFPE IHC staining?

    Thank you very much !

    Read More

    Abcam community

    Verified customer

    Asked on Aug 07 2012

    Answer

    Thank you for contacting us.

    We haven't tested this antibody in paraffin embedded tissue sections (IHC-P), only frozen sections from mouse. However, IL10 is produced and secreted by a variety of cell lines, including T-cells, macrophages, mast cells and other cell types. Any tissue that has a high concentration of these cells would serve as a good positive control. Other IL10 antibodies have been used to stain colon and hippocampus, so theoretically the IL10 protein should be found in these tissues regardless of which antibody is used.

    https://www.abcam.com/IL-10-antibody-ab34843.html&tab=abreviews&intabreviewid=16485

    https://www.abcam.com/IL10-antibody-2G101H7-ab25073.html

    And although it is for human IL10, you should check out the IHC staining from the Human Protein Atlas for IL10.

    http://www.proteinatlas.org/ENSG00000136634/normal

    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

    Read More

    Abcam Scientific Support

    Answered on Aug 07 2012

    Question

    I am using the Abcam IL10 [JES5-2A5] antibody. I am trying to optimize my Western Blot by having a positive control lane containing recombinant IL10. I see on the product info sheet that you have a figure showing exactly this (IL10 band from recombinant IL10). I was wondering if you could please advise me know on how much recombinant protein I should use in a positive control lane?

    Read More

    Abcam community

    Verified customer

    Asked on May 09 2012

    Answer

    We typically load 10 - 20 nanograms of purified recombinant protein in gels for western blots. I have contacted the laboratory to find out the exact amount that was used for the blot on the datasheet but I expect it will be within that range.

    Read More

    Abcam Scientific Support

    Answered on May 09 2012

    Question

    Could you please recommend me an isotype control for ab33471? I am using this antibody for WB and immunofluorescence of frozen tissues.

    Read More

    Abcam community

    Verified customer

    Asked on May 07 2012

    Answer

    Thank you for contacting us.

    We recommend ab18412.

    Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on May 07 2012

    Question

     Thank you for your prompt response to the inquiry about antibody ab33471 (IL-10).  We have had success with Abcam antibodies in the past and will continue to order from Abcam.  Unfortunately this antibody has not proven reliable based upon the datasheet. We have been trying to quantify IL-10 using Western blotting techniques.  From your datasheet a 17kDa band should be visualized with a 1:1000 dilution of the antibody.  Unfortunately, we are unable to resolve any bands using this and higher concentrations. In response to your request for information regarding the antibody, I have provided the following informations: Order number - ***   Samples used:  Mouse colon tissue lysates.  Snap frozen in liquid nitrogen, then processed in lysis buffer and homogenized. Blocking solution:  5% milk solution in Tris-NaCL-Tween 20 solution. Primary Antibody dilution ranged from 1:1000 to 1:250. HRP conjugated secondary antibody dilution ranged from 1:5000 to 1:1000. Detection System:  Thermo Fisher West Pico Chemiluminescent and Thermo Fisher West Dura Chemiluminescent Substrate Attached is a photo of a western blot with mouse tissue lysates probed with Abcam IL-10 (ab33471) at 1:500. If this information meets your criteria, please let me know how to proceed for a refund. Thanks you,    

    Read More

    Abcam community

    Verified customer

    Asked on Nov 23 2011

    Answer

    Thank you for your reply and for sending your protocol information and image. We do guarantee ab33471 to work in Western blot with mouse lysates, so we are happy to issue you a refund for this purchase. I have requested the refund to our accounting department, and this purchase will be refunded back to your credit card. If you have any questions about the status of this refund, please contact us.credits@abcam.com and refer to credit note ***. Please let me know if you have any questions, or if there is anything else that we can do for you.

    Read More

    Abcam Scientific Support

    Answered on Nov 23 2011

    1-10 of 13 Abreviews or Q&A

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