Anti-IL-6 antibody (ab6672)

Thank you for contacting us.

We recommend a 2:1 ratio of peptide to antibody. I have attached our protocol for your review.

Please let me know when are able to find the PO or Abcam order number for this purchase. Alternatively, if you know the date ordered and name of the purchaser I may be able to find this information as well.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for your patience as it has taken me longer than anticipated to gather the information necessary to help you solve the issue of non specific binding which you have been experiencing when using ab6672.

I have been able to add the immunizing peptide for ab6672 to our catalog with the designation ab133194. Using this product as a blocking peptide will help to determine which band or staining is specific. Briefly, the antibody (ab6672) is neutralized by incubation with its immunizing peptide (ab133194). The antibody that is bound to the blocking peptide is no longer available to bind to the epitope present in the protein on the western blot. The neutralized antibody is then used side-by-side with the antibody alone, and the results are compared. By comparing the staining from the blocked antibody versus the antibody alone, you can see which staining is specific as the specific staining will be absent from the western blot performed with the neutralized antibody.

If you would like to use this in order to determine specific bands for your experiments using ab6672 I would be happy to send you this peptide free of charge. To do this, would you please provide the Abcam confirmation number or PO used to purchase ab6672?

Please let me know how you would like to proceed and if you have any further questions.

Thank you for contacting us. I am sorry that you have been having difficulty conjugating ab6672 to fluorescent beads.

This antibody is supplied as whole antiserum, and it is therefore not unusual to see some red or pink discoloration. We will continue to guarantee the antibody for use in the tested applications listed on our datasheet: WB, ELISA, IHC-P, RIA, IP, IHC-Fr, Neutralising, ICC/IF.

We have not validated this antibody for use in flow cytometry, but it is possible it may be suitable for use in that application. Could you please provide some additional details about what type of beads you are using and what the conjugation chemistry is? Without knowing further detail, my main theory would be that the beads are conjugating to the normal rabbit IgGs present in this antibody. Whole antiserum will contain around 10 mg/mL of normal rabbit antibodies in addition to the ˜0.5 mg/mL of specific antibodies directed against IL6. When using our EasyLink conjugation kits, we sometimes observe a reduction in specific signal when the conjugate binds to other proteins in the solution.

I hope this helps. I will be more than happy to help you further once I have some additional details about the type of conjugation you are doing.

Vielen Dank für Ihre Antwortund dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

Es tut mir leid, dass Sie Probleme mit unseren Antikörpern hatten.

Um Ihr Problem zu lösen, möchte ich die folgenden Vorschläge zu Veränderung Ihres Protokolls machen:

Es ist sehr unwahrscheinlich, dass drei Antikörper gegen verschieden Zielprotein nicht funktionieren. Daher bin ich zuversichtlich, dass mit Protokolloptimierung eine Besserung der Ergebnisse erreicht werden kann.

1.) Der gleiche sekundäre Antikörper wird fier alle drei primären Antikörper benutzt. Funktioniert dieser mit anderen primären? Ich schlage vor einen anderen sekundären auszuprobieren, am besten auch mit einem anderen Fluorochrom um Probleme mit dem Mikroskop auszuschließen.

2.) Für die meisten Zielproteine ist Citrate Puffer als Antigendemaskierung suboptimal. ich schlage deshalb vor EDTA, pH 9 auszuprobieren. ich habe mehr Infomaterial als Attachement angehängt.

3.) Da es sich um sekretierte Proteine handelt ist eine Permeabilisierung nicht unbedingt nötig. Ich schlage vor allem vor, nicht Triton und Tween in der Lösung für den sekundären Antikörper zu verwenden. So kann TBS 0.025% Triton als Puffer für die Antikörper verwendet werden.

4.) Bei zu wenig oder fehlendem Signal können die primären Antikörper auch nur in TBS 0.025% Triton inkubiert werden. Ich stimme ihnen zu, dass über Nacht bei 4C die beste Option ist.

Bitte lassen Sie mich wissen, ob ich Ihnen helfen konnte und zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Thank you for your inquiry.

I can confirm thatone of the imageson the datasheet shows rat knee positive control with induced arthritis.

The good news is that we do have plenty of Abreview where human samples were successfully used:

Pancreatic cancer , Shoulder- Bursa, head and neck tumor and liver.

Please follow this link to the datasheet and to review the Abreviews:

https://www.abcam.com/IL-6-antibody-ab6672.html (or use the following: https://www.abcam.com/IL-6-antibody-ab6672.html).

There are also many publications that haveused this antibody successful in human tissues. maybe they can offer some help when deciding on a suitable positive control?

https://www.abcam.com/IL6-antibody-ab6672-references.html

Which application are you using? We will be happy to provide technical help if it still proves difficult to work with this antibody after using a positive control.

I hopethis information is helpful. Please feel free tocontact me again withany further questions.

Thank you for contacting Abcam.

I understand your concerns about the appearance of ab6672 that you received. This IL6 antibody is provided as whole antiserum and as such it is not unusual that the liquid sometimes might appear bright red. This is due to haemolysis of erythrocytes which can occur during the sterile filtration of the serum. To our experience this does not affect the performance of the antibody.

Under out Abpromise we guarantee this product to work as stated on the datasheet and if you experience any problems with it, we will be happy to help resolve these issues. In the event that we cannot solve the issues you are having, then we would be happy to either replace or refund.

If there is anything else I can help you with, please let me know.

Thank you for contacting us and reporting the results you have obtained with Anti-IL6 antibody (ab6672).


The bands observed at ˜55 kDa in the western blot presented on the datasheet of ab6672 has not been fully characterised. It may be that it is a multimer of IL-6 or some non-specific binding. It may be with further optimisation of blocking conditions this could be reduced further.


If you would like I can have a look at your blot and protocol to see if I am able to suggest any modifications which may improve the specificity. If you would be interested in me doing this, please fill out the questionnaireI have attached to this email.


Otherwise, if you have any further questions please do not hesitate to contact us again.

Thank you for contacting us.

Unfortunately we do not provide positive controls with our antibodies.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

Thank you very much for contacting us.

I understand your concerns about the appearance of the vial of ab6672 you received. I would like to reassure you that we monitor customer feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this lot number. This IL6 antibody is provided as whole antiserum and as such it is not unusual that the liquid sometimes might appear bright red. This is due to haemolysis of erythrocytes which can occur during the sterile filtration of the serum. To our experience this does not affect the performance of the antibody.

We guarantee this antibody to work in the applications stated on the datasheet, including immunohistochemistry, and recommended dilutions for each application are also listed on the datasheet. Although you may have established the protocol already, we would like to recommend to adjust dilutions appropriately from the results obtained. This fine-tuning or optimisation is common scientific practise for any new antibody lot, as due to the nature of polyclonal antibodies the same affinity cannot be guaranteed for different antibody production lots.

Please be reassured that in the event that the product is not functioning as cited on the datasheet, we will be pleased to provide a credit note, free of charge replacement or refund, as you are covered by our Abpromise (www.abcam.com/abpromise).

I hope this information is helpful, thank you for your cooperation. I would like to wish you good luck with their research. The technical team is always at your service, should you require further expert advice.

Thank you for contacting Abcam regarding ab6672.

I have confirmed with the laboratory that unfortunately we do not have any additional concentration information for this antibody. As it is an unpurified whole antiserum, we can only estimate the concentration is 1-5mg/ml.

I hope this information is helpful and I apologize for any inconvenience. Please do not hesitate to contact us if you have any additional questions.