Overview

  • Product name
    Human IL-6 ELISA Kit High Sensitivity
    See all IL-6 kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 4.4%
    Inter-assay
    Sample n Mean SD CV%
    Sample 1 18 25pg/ml 3 12.2%
    Sample 2 18 12pg/ml 1 8.7%
    Sample 3 18 6pg/ml 1 13.7%
    Sample 4 18 41pg/ml 2 5.9%
    Sample 5 18 31pg/ml 1 4.9%
  • Sample type
    Cell culture supernatant, Serum, Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 0.8 pg/ml
  • Range
    1.56 pg/ml - 50 pg/ml
  • Recovery

    93 %

  • Assay time
    3h 45m
  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Abcam’s IL-6 Human High Sensitivity in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-6 in supernatants, buffered solutions, serum, plasma and other body fluids. This assay will recognise both natural and recombinant Human IL-6.


    A monoclonal antibody specific for IL-6 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known IL-6 concentrations, control specimens or unknowns are pipetted into these wells. During the first incubation, the standards or samples and a biotinylated monoclonal antibody specific for IL-6 are simultaneously incubated. After washing, the enzyme Streptavidin-HRP, that binds the biotinylated antibody is added, incubated and washed. A TMB substrate solution is added which acts on the bound enzyme to induce a colored reaction product. The intensity of this colored product is directly proportional to the concentration of IL-6 present in the samples. 


    This kit will recognize both endogenous and recombinant Human IL-6.


    Get results in 90 minutes with Human IL-6 ELISA Kit (ab178013) from our SimpleStep ELISA® range.


     

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components Identifier 1 x 96 tests 2 x 96 tests
    10X Standard Diluent Buffer Black 1 x 25ml 1 x 25ml
    200X Wash Buffer White 1 x 10ml 2 x 10ml
    Biotinylated Antibody Diluent Red 1 x 7.5ml 1 x 13ml
    Biotinylated anti-IL-6 Red 1 x 0.4ml 2 x 0.4ml
    Chromogen TMB Substrate Solution 1 x 11ml 1 x 24ml
    HRP Diluent Red 1 x 23ml 1 x 23ml
    IL-6 HS Control Silver 2 vials 4 vials
    IL-6 Microplate (12 x 8 well strips) 1 unit 2 units
    IL-6 HS Standard (Lyophilized) Yellow 2 vials 4 vials
    Plastic Plate Covers 1 x 2 units 1 x 4 units
    Standard Diluent (Serum) 1 x 7ml 2 x 7ml
    Stop Reagent Black 1 x 11ml 2 x 11ml
    Streptavidin-HRP 2 x 5µl 4 x 5µl
  • Research areas
  • 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
  • Alternative names
    • Interleukin BSF 2
    • B cell differentiation factor
    • B cell stimulatory factor 2
    • B-cell stimulatory factor 2
    • BSF 2
    • BSF-2
    • BSF2
    • CDF
    • CTL differentiation factor
    • Hepatocyte stimulatory factor
    • HGF
    • HSF
    • Hybridoma growth factor
    • Hybridoma growth factor Interferon beta-2
    • IFN-beta-2
    • IFNB2
    • IL 6
    • IL-6
    • IL6
    • IL6_HUMAN
    • Interferon beta 2
    • Interferon beta-2
    • Interleukin 6
    • Interleukin 6 (interferon beta 2)
    • Interleukin BSF 2
    • Interleukin-6
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab46042 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • IL-6 was measured in cell culture medium from U937 control cells, or cells stimulated with LPS (1 �g x mL-1; 6 h) or PMA (10 ng x mL-1; 24 h) and LPS (background signal subtracted) (duplicates +/- SD).
  • Representative Standard Curve using ab46042

Protocols

References

This product has been referenced in:
  • Yang Z  et al. miR-143-3p regulates cell proliferation and apoptosis by targeting IGF1R and IGFBP5 and regulating the Ras/p38 MAPK signaling pathway in rheumatoid arthritis. Exp Ther Med 15:3781-3790 (2018). Read more (PubMed: 29581736) »
  • Mayr HL  et al. Randomization to 6-month Mediterranean diet compared with a low-fat diet leads to improvement in Dietary Inflammatory Index scores in patients with coronary heart disease: the AUSMED Heart Trial. Nutr Res 55:94-107 (2018). Read more (PubMed: 29754829) »
See all 18 Publications for this product

Customer reviews and Q&As

1-10 of 13 Q&A

Answer

1. The Biotinylated Antibody is provided in liquid form, it is stable at 4°C
2. Standards and Controls are provided lyophilized and should be reconstituted either by the volume stated on the vial or in the Protocol Booklet. They should be used directly once reconstituted and can't be stored.


3. The Standard and Control are both made with recombinant IL-6, only the concentration differs.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1202121.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that our source for this kit is kindly providing this replacement, and so we are waiting for this to come to us before we can send it to you. I do not anticipate this will take too long. They are providing a different lot.

The free of charge replacement kit will also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let me know.

I wish you the best of luck with your research.

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Answer

Thank you for your message and for providing this further information.

I have reviewed this enquiry and discussed it with the originator of this kit. Regrettably, it does seem the second kit is not providing the sensitive results you require. I appreciate the time you have spent on these experiments and would be pleased to arrange a free of charge replacement or credit note in compensation for the second kit which did not work so well.

I look forward to hearing from you with details of how you would like to proceed.

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Answer

Thank you for taking the time to contact us. Your message has been forwarded to the scientific support team. I am sorry to hear you have had difficulty obtaining satisfactory results from the second kit you received.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab46042 is tested and covered by our 6 month guarantee for ELISA on human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, In order to help me investigate, I would appreciate if you can confirm some further details:

1. Please provide the order number and date of purchase

2. What type of samples have been tested, from which species?

3. What were the kit storage conditions?

4. Please describe the problem in further detail. I would appreciate if you are able to provide some data, particularly the standard curves from each kit, which will help me to assess the results.

Thank you for your cooperation. Please do not hesitate to contact me again with the further requested details and I hope we can resolve this case for you as soon as possible.

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Answer

Thank you for your inquiry.
Do you have the catalogue numbers of the kits you're inquiring about? We have 7 different IL6 Human ELISA kits in the catalogue: ab46044, ab46042, ab46027, ab46041, ab100572, ab100573 and ab108867.
The IL6 High Sensitivity kits (ab46044 and ab46042) are the same, just one is 1 x 96 tests and one is 2 x 96 tests. These are high sensitivity at < 0.8 pg/ml and is in sandwich format and works with cell culture supernatant, Serum, and Plasma.
The ab46027 works with Cell culture supernatant, Serum, and Plasma as well in a sandwich format, but the sensitivity is < 2 pg/ml, but greater recovery.
The ab100572 has a broader range 1.37 pg/ml - 1000 pg/ml but < 3 pg/ml sensitivity. This works in Cell culture supernatant, Serum, and Plasma as well.
The ab100573 is the same as ab100572, except that ab100573 is recommended for cell culture extracts and tissue extracts.
The ab1008867 works with works with Cell culture supernatant, Serum, and Plasma as well in a sandwich format but is the least sensitive (8 pg/ml).
If you purchase multiple kits, you will only pay 1 shipping charge. If an item on your order is out of stock, your order will be on hold until it is ready to ship. However, you can choose to "split" your order to receive the in-stock item the next day, and the out of stock item when it comes in, but you would have to pay for both shipping charges in that case.
I hope this information helps. Please contact us with any other questions.

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Question
Answer

Thank you for contacting us.

The lab let me know the following:
Saliva is suitable for thisELISA kit, and the regular protocol would apply. There is no need to pre-treat the samples before assaying.

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

Thank you for confirming those details.

I have noticed that the results that you provided me with, there is a difference between the positive and negative samples by eye. How has your spectrophotometer been set up to measure this? At what wavelength have you been reading?

How long did you leave the colour to develop with the TMB substrate?

Also, I am not sure of the dilutions you describe. What was the control IL-6HS and standard IL-6HS?

You were provided with a vial of control, was this reconstituted in 1 mL of human serum diluent, then diluted further?

I am sorry for all the questions but I am trying to isolate which step may be contributing to these problems. I am also waiting on some further feedback from the lab and hopefully they will have some further suggestions to make.

I am sorry for the delay and the inconvenience this may be causing you.

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Answer

Thank you for your information.

I am sorry for the delay in getting back to you. I think it would be very helpful for us to understand exactly how the standard samples and streptavidin HRP was prepared.

As you say, this kit has been extensively optimised to allow the customer to simply use it without performing extra optimisation. We have had no other reports of the kit not working as expected and its quite a popular kit. I would therefore really like to understand in detail how the solutions were made up in order to try and help you as best I can.

Can you therefore state, in uL, how each solution used was made up and diluted?

I really appreciate your help and hope that I can advise you as to how to get the best results out of this kit.

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Question

Hi
below are the answers to all questions.
1. You mentioned thatyou used 3 strips. Were all of these using the standard provided with the kit? What results did you obtain with the other 2 strips? For the time , i did run only 3 strips ( 2 forstandardsand one strip for 4samples), there are the result which i had..
strip 1(standards)
strip 2 (standards)
strip 3 (samples in duplicate)
AND THE SECOND TIME I DID RUN ONLY 1 STRIP ( THE STANDARD ONLY ), i have emailed you the results that i had for this time.
2. Were all reagents allowed to come to room temperature before use? YES, all reagents allowed to room temperature before use.
3.Exactly how was the standard prepared and diluted? What diluent was used? because i used a human sample (serum) so i used the standerd diluent : human serum which is ready to use ( no need to dilute ).
4. When washing, was the excess liquid removed by aspiration or tapping plate on absorbent paper? Tapping the plate onabsorbentpaper
5. Was the TMB solution protected from the light? Yes it was in the kit's box, i only took it out of the box before use it and i did protect the plate from the light by wrapping it withaluminumfoil.
6. What colour was the TMB solution before adding to the wells?colorless
7. How was the biotinylated anti-IL-6 prepared? according to the no of stripes i need, so i did add 60µ of Biotinylated Antibody concentrate and 1590µ
of Biotinylated Antibody diluent in a clean glass tube.
8. How was the Streptavidin-HRP prepared? i add 200µ from Diluent and 2µ from HRP.
9. Were the strips covered during the 3 hour incubation of the biotinylated anti-IL6? YES they were covered.
10. Was the HRP solution prepared directly prior to use? yes it was prepared immediately before use.
11.At whatwavelength was the absorbancemeasured? What kind of reader have you been using? at 450 nm
12. The wash steps you refer to, after which steps werethese performed and exactly how did you perform them? exactly as its written in the manual protocol, by tapping the pale on absorbent paper.
thanks

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Answer

Thank you for confirming those details.

I am not entirely sure why you are seeing such low readings. It is likely to do either with the standard itself or with the detection (antibody and HRP conjugate). What I would like to do if you wouldn't mind is provide you with a replacement kit free of charge to see if the kit has been damaged in any way. This is very uncommon but as I can see no problem in how you have been using the kit I would like to rule this out before trying some optimisation or the protocol.

If you would like for me to do this could you please provide me with the order number (or delivery address and approximate date of delivery) and I will have this arranged as soon as possible.

I look forward to receiving your reply.

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Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

In order to understand what may be contributing to the low readings and potentially offer solutionscould you please answer the following questions:

1. You mentioned thatyou used 3 strips. Were all of these using the standard provided with the kit? What results did you obtain with the other 2 strips?

2. Were all reagents allowed to come to room temperature before use?

3.Exactly how was the standard prepared and diluted? What diluent was used?

4. When washing, was the excess liquid removed by aspiration or tapping plate on absorbent paper?

5. Was the TMB solution protected from the light?

6. What colour was the TMB solution before adding to the wells?

7. How was the biotinylated anti-IL-6 prepared?

8. How was the Streptavidin-HRP prepared?

9. Were the strips covered during the 3 hour incubation of the biotinylated anti-IL6?

10. Was the HRP solution prepared directly prior to use?

11.At whatwavelength was the absorbancemeasured? What kind of reader have you been using?

12. The wash steps you refer to, after which steps werethese performed and exactly how did you perform them?

I am sorry for all the questions but I am hoping to be able to understand what may be contributing to the low readings you have been observing with the standard. If possible could you also provide me with the order number or the approximate date of delivery and the delivery address?

I thank you for your cooperation and I look forward to receiving your reply.

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1-10 of 13 Q&A

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