Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab9324 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
IHC-Fr 1/1000.
ICC/IF Use at an assay dependent concentration.
Neutralising Use at an assay dependent concentration. To yield one-half maximal inhibition [ND50] of the biological activity of hIL-6 (0.5 ng/ml), a concentration of 19.0-23.0 ug/ml of this antibody is required.
Sandwich ELISA Use at an assay dependent concentration. In a sandwich ELISA (assuming 100µl/well), a concentration of 4.0-8.0 µg/ml of this antibody will detect at least 500 pg/ml of recombinant human IL-6 when used with Rabbit polyclonal to IL6 (Biotin) (ab84251) as the detection antibody at a concentration of approximately 0.5-1.0 µg/ml.
IHC-P Use a concentration of 0.125 µg/ml.
WB Use a concentration of 0.2 - 0.4 µg/ml. Used in conjunction with compatible secondary reagents, the detection limit for recombinant human IL-6 is 0.5-1.0 ng/lane, under reducing or non-reducing conditions.

Target

  • Function
    Cytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Plays an essential role in the final differentiation of B-cells into Ig-secreting cells Involved in lymphocyte and monocyte differentiation. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation Acts on B-cells, T-cells, hepatocytes, hematopoeitic progenitor cells and cells of the CNS. Also acts as a myokine. It is discharged into the bloodstream after muscle contraction and acts to increase the breakdown of fats and to improve insulin resistance.
  • Involvement in disease
    Genetic variations in IL6 are associated with susceptibility to rheumatoid arthritis systemic juvenile (RASJ) [MIM:604302]. An inflammatory articular disorder with systemic-onset beginning before the age of 16. It represents a subgroup of juvenile arthritis associated with severe extraarticular features and occasionally fatal complications. During active phases of the disorder, patients display a typical daily spiking fever, an evanescent macular rash, lymphadenopathy, hepatosplenomegaly, serositis, myalgia and arthritis.
    Note=A IL6 promoter polymorphism is associated with a lifetime risk of development of Kaposi sarcoma in HIV-infected men.
  • Sequence similarities
    Belongs to the IL-6 superfamily.
  • Post-translational
    modifications
    N- and O-glycosylated.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • Interleukin BSF 2 antibody
    • B cell differentiation factor antibody
    • B cell stimulatory factor 2 antibody
    • B-cell stimulatory factor 2 antibody
    • BSF 2 antibody
    • BSF-2 antibody
    • BSF2 antibody
    • CDF antibody
    • CTL differentiation factor antibody
    • Hepatocyte stimulatory factor antibody
    • HGF antibody
    • HSF antibody
    • Hybridoma growth factor antibody
    • Hybridoma growth factor Interferon beta-2 antibody
    • IFN-beta-2 antibody
    • IFNB2 antibody
    • IL 6 antibody
    • IL-6 antibody
    • IL6 antibody
    • IL6_HUMAN antibody
    • Interferon beta 2 antibody
    • Interferon beta-2 antibody
    • Interleukin 6 antibody
    • Interleukin 6 (interferon beta 2) antibody
    • Interleukin BSF 2 antibody
    • Interleukin-6 antibody
    see all

Images

  • IHC image of IL6 staining in human spleen formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9324, 1in250 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-IL-6 antibody (ab9324) at 1/2000 dilution

    Lane 1 : Human Spleen
    Lane 2 : Human Lung
    Lane 3 : Rat Spleen
    Lane 4 : Rat Lung

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : 800CW Goat Anti-Mouse IgG at 1/10000 dilution

    Performed under reducing conditions.

    Observed band size: 17,25 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 50 kDa (possible multimer)



    This antibody was raised against an immunogen that is predicted to recognize the glycosylated form of IL6 as well as the IL6delta4 splice variant. The predicted molecular weights are 25 kDa and 17 kDa respectively. The band observed at 50 kDa may represent multimers of IL6 as reported in the literature.

  • Immunohistochemical analysis of PFA-fixed frozen murine neural tissue sections, labelling IL6 with ab9324 at a dilution of 1/1000 incubated for 24 hours at 4°C in 0.1M PBST with 10% donkey serum. Permeabilization was with 0.1M PBS and 3% Triton X. Blocking was with 10% serum incubated for 1 hour at 24°C. Secondary was a mouse monoclonal Alexa Fluor® 568 conjugate at 1/1000.

    See Abreview

  • ab9324 staining IL6 in human cervical squamous cell carcinoma tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections). Tissue underwent heat mediated antigen retrieval in sodium citrate buffer (pH 6.0). The primary antibody was used at 0.125 ug/ml and incubated with sample at 4°C overnight. A HRP-labeled polymer detection system was used with a DAB chromogen.

  • ICC/IF image of IL6 staining (ab9324) in rat primary microglial cell culture. The sections were incubated in 10% serum to block non-specific protein-protein interactions and in 0.3% triton X in 0.1% PBS for 1h to permeabilise the cells. The sections were then incubated with ab9324 (1:1000) overnight at +4°C, followed by Alexa 568 conjugated secondary antibody. Red staining of the cytosol was observed.

    See Abreview

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) »
  • Liu Z  et al. Number 2 Feibi Recipe Reduces PM2.5-Induced Lung Injury in Rats. Evid Based Complement Alternat Med 2018:3674145 (2018). Read more (PubMed: 29541141) »
See all 45 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (retina)
Permeabilization
No
Specification
retina
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Paraformaldehyde

Mr. Tamas Bellak

Verified customer

Submitted Mar 27 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Muller Cell Line MIO-M1)
Permeabilization
Yes - 0.1% TritonX-100
Specification
Muller Cell Line MIO-M1
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 01 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse 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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: rt°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Tris, 1mM EDTA pH 9
Sample
Monkey Tissue sections (retina)
Specification
retina
Permeabilization
No
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 10 2015

Abreviews
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (4-15% SDS-PAGE)
Sample
Monkey Tissue lysate - whole (macaque retina)
Specification
macaque retina
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 10 2015

Answer

All of these antibodies recognise cytokines which are present in inflammed tissue therefore any inflammed tissue would be a good positive control.

In our catalogue, we have several cancer slides which would be appropriate:
tonsil: (ab5146)
breast (ab4701)
pancreatic (ab5036)
stomach (ab5115)
colon (ab4702)

Alternatively, spleen tissues seem to be a very good positive control for these cytokines and are used on several of our other datasheets.

I would suggest using ab4372 if you are working on human samples, ab4613 if you are working on mouse samples or ab4624 if you are working on rat samples.

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Answer



Any products that require an MSDS form will have a link on the product's online datasheet to the data. It will be a PDF document link labeled 'SDS'. Please see attached image for example of what to look for on the datasheet. If no 'SDS' link is available than the product does not contain any components that require an SDS.

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Answer

DISCOUNT CODES:
***

Expiration date: November 26, 2012

These codes will each give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for each antibody and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Eachcode will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you very much for your call today and for your interest in our IL6 and IL8 antibodies.

I've looked over the rest of our IL6 antibodies, and we unfortunately don't have any that Istrongly expect to cross-react with rabbit samples. The mouse monoclonal ab9324 could cross-react, as it does react with rat samples which are also dissimilar to the immunogen human sequence.

We also have a rabbit polyclonal antibody, ab6672, which was raised against the full-length human protein and may have a higher chance of cross-reaction since it is polyclonal. However if you willbe staining tissue with expected endogenous IgG molecules (i.e. spleen or tonsil) thentherecan behigh backgroundfrom reaction between the tissue and the anti-rabbit secondary antibody. I'm not sure which IL6 antibodywill be the better option for you to try, but if you would like to try either of these, we do have a testing discount program (see details below for ab34100) that you may be interested in.

Since the IL8 antibody ab34100 has not been tested in IHC-P,I can offer a discount off a future purchase if you buy ab34100 now, test it in IHC-P,and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would beworth 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab34100 in IHC-P. I will then send a discount code. This code must be issued before purchasing ab34100 so please wait for my reply before ordering.

2. Purchase ab34100 either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-P.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab34100 turns out to be unsuitable for IHC-P, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us.

The anti-IL6 antibody ab9324 is a mouse monoclonal IgG2a. Therefore, you will need to use a mouse monoclonal IgG2a isotype control.

Therefore, you should use the mouse control IgG2a ab18413 and NOT the rabbit IgG ab37415.

To learn more about isotype controls, please see the link below:

https://www.abcam.com/index.html?pageconfig=technicalfaqs#5

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

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1-10 of 18 Abreviews or Q&A

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

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