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: WB, IHC-Fr, ELISAmore 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.

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: 7.2
    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

Applications

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
WB Use at an assay dependent concentration. Detects a band of approximately 17 kDa.

We recommend incubating the membrane overnight in primary antibody.

IHC-Fr Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.

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

  • All lanes : Anti-IL-10 antibody [JES5-2A5] (ab33471) at 1/500 dilution

    Lane 1 : Mouse liver whole cell lysate (treated with LPS 5mg/kg)
    Lane 2 : Mouse hepatocytes (Treated with LPS 100 ng/ml)

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : Goat anti-rat (HRP) at 1/1000 dilution

    Developed using the ECL technique.

    Observed band size: 17 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 30 kDa (possible non-specific binding)


    Exposure time: 45 seconds


    Blocked with 5% milk for 18 hours at 4°C.

    See Abreview

  • Immunohistochemistry (Frozen sections) analysis of rat brain tissue sections labeling IL-10 with ab33471 at 1/1000 dilution. The tissue was fixed with paraformaldehyde and permeabilized with 0.1 M PBS with 3% Triton X. The tissue was blocked with 10% serum for 1 hour at 24°C, followed by incubation with Anti-IL-10 antibody [JES5-2A5] (ab33471) in 0.1M PBST with 10% donkey serum for 24 hours at 4°C. Alexa Fluor 488® secondary antibody was used at 1/1000 dilution. The IL-10 was visible in the M2 macrophages in the choroid plexus.  Nucleus was stained with DAPI.

References

This product has been referenced in:

  • Nakayama T  et al. Regeneration-associated cell transplantation contributes to tissue recovery in mice with acute ischemic stroke. PLoS One 14:e0210198 (2019). Read more (PubMed: 30682162) »
  • Wang G  et al. Age-Associated Dopaminergic Neuron Loss and Midbrain Glia Cell Phenotypic Polarization. Neuroscience 415:89-96 (2019). Read more (PubMed: 31325563) »
See all 20 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

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

Dr. WeiHua Wu

Verified customer

Submitted Mar 25 2019

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

Dr. WeiHua Wu

Verified customer

Submitted Mar 25 2019

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

Abcam user community

Verified customer

Submitted Jan 23 2017

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 Jun 25 2015

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

Abcam user community

Verified customer

Submitted May 13 2014

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

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/index.html?datasheet=34843&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

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

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

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.

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1-10 of 13 Abreviews or Q&A

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