Overview

  • Product name
  • Description
    Rabbit polyclonal to IL-10
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ELISA, Neutralising, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment corresponding to Rat IL-10 aa 19-178.
    (Peptide available as ab9970)

Properties

  • Form
    Lyophilised:Reconstitute with 200µl of sterile water. Please note that if you receive this product in liquid form it has already been reconstituted as described and no further reconstitution is necessary.
  • 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
    PBS, pH 7.4, no preservative, sterile filtered
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Light chain type
    unknown
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab9969 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 a concentration of 0.1 - 0.2 µg/ml. Can be blocked with Recombinant rat IL-10 protein (ab9970).
ELISA Use a concentration of 0.5 µg/ml.
Neutralising Use at an assay dependent dilution. To yield one-half maximal inhibition [ND50] of the biological activity of rIL-10 (30.0 ng/ml), a concentration of 0.45 - 0.8 µg/ml of this antibody is required.
ICC/IF 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

  • ab9969 staining IL10 in murine immortalised bone marrow-derived macrophage by Immunocytochemistry/ Immunofluorescence. The cells were fixed in paraformaldehyde, permeabilised in 0.01% Triton X-100 and then blocked using 5% serum for 1 hour at 20°C. Samples were then incubated with primary antibody at 1/200 for 1 hour at 20°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 568 (red) used at a 1/500 dilution.

    See Abreview

  • ab9969 staining IL10 in RAW 246.7 cells treated with spermidine hydrochloride (ab120057), by ICC/IF. Increase in IL10 expression correlates with increased concentration of spermidine hydrochloride, as described in literature.
    The cells were incubated at 37°C for 24 hour in media containing different concentrations of ab120057 (spermidine hydrochloride) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab9969 (1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • ab9969 staining nucleolin in Raw 264.7 cells treated with spermine (ab120051), by ICC/IF. Increase in IL10 expression correlates with increased concentration of spermine, as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab120051) (spermine) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab9969 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

References

This product has been referenced in:
  • Xiang Q  et al. ISL1 overexpression enhances the survival of transplanted human mesenchymal stem cells in a murine myocardial infarction model. Stem Cell Res Ther 9:51 (2018). Read more (PubMed: 29482621) »
  • Tu R  et al. Soluble epoxide hydrolase inhibition decreases reperfusion injury after focal cerebral ischemia. Sci Rep 8:5279 (2018). Read more (PubMed: 29588470) »
See all 27 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (vessel)
Permeabilization
Yes - polysorbate 20
Specification
vessel
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 06 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (brain)
Specification
brain
Blocking step
1% Goat serum 1% BSA 5% NFDM in 1X PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Feb 17 2014

Answer

Vielen Dank für Ihre Anfrage.

Ich habe vier primäre AK für Sie herausgesucht, die alle in einer relativ hohen Verdünnung eingesetzt werden können, so dass sie für eine Weile reichen sollten. Das Sie alle Proben nun damit verarbeitet bekommen kann ich leider nicht versprechen, da die Antikörper ja auch in den meisten Fällen ein paar Optimierungsexperimente benötigen.

https://www.abcam.com/index.html?datasheet=9739 (or use the following: https://www.abcam.com/index.html?datasheet=9739).

https://www.abcam.com/index.html?datasheet=9969 (or use the following: https://www.abcam.com/index.html?datasheet=9969).

https://www.abcam.com/index.html?datasheet=13243 (or use the following: https://www.abcam.com/index.html?datasheet=13243).

https://www.abcam.com/index.html?datasheet=83339 (or use the following: https://www.abcam.com/index.html?datasheet=83339).


Als Sekundär-AK würde ich Ihnen den ab97069 Goat anti-Rabbit IgG H&L (HRP) secondary antibody empfehlen, da dieser bis zu einer Verdünnung von 1/50.000 eingestzt werden kann.



https://www.abcam.com/index.html?datasheet=97069 (or use the following: https://www.abcam.com/index.html?datasheet=97069).


Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere technische Fragen haben. Falls Sie die Produkte Bestellen möchten oder mehr Informationen zB bezüglich des Preises wünschen, wenden Sie sich einfach an unseren Kundendienst.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

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Answer

Thank you very much for contacting us with your enquiry.

The antibody ab9969 may be suitable for use with paraffin-embedded tissue, however we can't say for sure until this is tested. For cases like this, in which you're interested in trying an antibody in an application that we haven't yet tested, we do have a testing discount program called AbTrial that you can participate in. As part of our AbTrial program, you would purchase ab9969 and test it in IHC-P, and then submit an Abreview with your data and protocol. In exchange, we will give you a discount code worth $337 (the cost of ab9969) off of a future purchase.

I am sorry that we don't have an antibody that has already been tested in your assay, but if you'd like more information about AbTrial or if you have any questions, please let me know and I'll be happy to help you further.

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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

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Answer

Thank you for contacting us. As an isotype control for ab9969, I would recommend either ab37415 or ab27478. I hope this helps, please let me know if you have any additional questions.

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Answer

Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results.

Can you tell me more about your IL10-/- mice? You said you checked by ELISA. Did you use this same antibody?

Have you tried any other fixatives? A customer had good results with formaldehyde.

https://www.abcam.com/index.html?datasheet=9969&tab=abreviews&intabreviewid=22906

If you do stick with the methanol fixation, I'd recommend omitting the triton-X premeabilization step then. Methanol fixes and permeabilizes at the same time, so the additional permeabilization may be disrupting the membrane too much.

You may also want to increase the blocking serum to 10% for 1 hr at RT and decrease the amount of primary antibody used.

Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

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Answer

Thank you for contacting us and your interest in our products.


As discussed over the phone ab8918isamouse monoclonaland canbe used with an anti-mouse IgG secondary antibody.


ab39626 and ab9969 are bothrabbit polyclonals andanti-rabbitIgG can be used in their detection.


If youdo find any non-specificitywith the anti-mouse secondary with your mouse sample (most likely a band at˜25 kDaand/or˜55 kDa)I would suggest looking at the followingprotocols:


https://www.abcam.com/index.html?pageconfig=resource&rid=11465

http://www.ihcworld.com/_protocols/general_IHC/immuno_mom.htm


Thesearefor IHC but should betransferable toWestern blotting. Alternatively I would suggest altering theprimary antibody to a non-mouseprimary.


I hope this information has beenof help. If you have any problems or require any further information please do not hesitate to ask.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (immortalised bone marrow-derived macrophage)
Specification
immortalised bone marrow-derived macrophage
Fixative
Paraformaldehyde
Permeabilization
Yes - .01% Triton X100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. James Harris

Verified customer

Submitted Jan 31 2011

Question
Answer

Thank you for your enquiry. The MW of rat IL10 is approximately 20 kDa. If you have any additional questions, please contact us again.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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