Overview

  • Product name
    Human IL-6 ELISA Kit
    See all IL-6 kits
  • Detection method
    Colorimetric
  • Sample type
    Cell culture extracts, Tissue Extracts
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 30 pg/ml
  • Range
    1.37 pg/ml - 1000 pg/ml
  • Recovery

    93 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 93.65 82% - 102%
    Tissue Extracts 92.78 81% - 101%

  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Abcam’s Human IL-6 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human IL-6 in cell lysates and tissue lysates.


    This assay employs an antibody specific for Human IL-6 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-6 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human IL-6 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IL-6 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    Get higher sensitivity in only 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 -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    2X Cell Lysate Buffer 1 x 5ml
    5X Assay Diluent 1 x 15ml
    5X Sample Diluent Buffer 1 x 10ml
    600X HRP-Streptavidin Concentrate 1 x 200µl
    Biotinylated anti-Human IL-6 (lyophilized) 2 vials
    IL-6 Microplate (12 x 8 wells) 1 unit
    Recombinant Human IL-6 Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • 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 ab100573 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

  • Representative standard curve using ab100573

Protocols

References

ab100573 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Our lab used this kit in order to assess IL-6 levels in the supernatant of cultured Jurkat T cells. We used the following protocol for our culture:
1. Frozen Jurkat cells were thawed and washed in warm complete media (1640 RPMI + 10% FBS + antibiotic).
2. Cells were then allowed to rest in warm media for an hour.
3. Cells were then supplied with new media alone or containing an antioxidant (30 mins). Cells were then washed with 1X DPBS and proceeded to next step.
4. Cells from the two conditions in "3" were then supplied with complete media or complete media + 162 nM PMA (24 hours).---work was done in a 6 well plate (using only 4 wells). Basically the conditions were: 1) +antioxidant/+PMA, 2)+antioxidant/-PMA, 3)-antioxidant/+PMA, 4)-antioxidant/-PMA
5. Cells and media were separated by centrifugation (1000g, 5 mins) and used in the ELISA kit (ab100573).
6. Cells (after wash) were lysed with the kits supplied lysis buffer and cell extract (supernatant) was separated by centrifugation----This is an area in the kit that needs more clarification. There is absolutely no guide on how much of the lysis buffer to use (volume:cell ratio), incubation time, recommended temp, centrifugation speed, etc, which made using the lysis buffer to get cell extract complete guess work.
7. Cells and cell culture supernatant (media) were assayed at various dilutions (1:4, 1:5, and 1:6 for cells and undiluted, 1:2, and 1:4 for culture supernatant).
The kit was very easy to use as we've used similar kits by abcam before, but we saw very low levels of IL-6 for the culture supernatant at all dilutions (in some cases undetectable) while we saw IL-6 levels for cells across all dilutions (though we did not see any real difference across groups).
Overall, it seems the kit works with cell lysate, but not cell culture supernatant.

Abcam user community

Verified customer

Submitted Mar 09 2016

Question
Answer

Thank you for contacting us.


The preparation for tissue using this kit is as follows.

2X lysis buffer is provided in the kit, and a protease inhibitor cocktail is recommended.

Choices of the method for lysis and homogenization include glass-bead “smash,” douncing,

freeze-thaw, sonication and crushing frozen tissue with a mortar and pestle, or even a

combination of these. There is no one “best method” for all sample types, but some are better

than others for some sample types. Your choice of method should be made following a brief

search of the literature to see how samples similar to yours have been prepared in previous

investigations.

After homogenization, spin down the lysates to remove cell/tissue debris (5 min @ 10000 x g

or 10 min @ 5000 x g) and save the supernatant. Lysates should be frozen as soon as

possible, and stored at -20°C (or -80°C, if possible). Spin them again before incubating with

the antibody array. Determine the protein concentration of your lysates (for example the

bicinchoninic acid (BCA) assay) and normalize the volume of each sample used to deliver the

same amount of total protein for each assay.

Since different cells and tissues may contain different amounts of protein, as starting point, we

suggest using 500 uL of lysis buffer per 1x106 cells or 10 mg tissue. You may have to adjust

this based upon your results. Your target for total protein concentration of the homogenate

should be at least 1000 ug/mL, but 2000 ug/mL would be better.




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

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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 your patience.

I've got great news - we have sourced the standards for these kits to be purchased separately! The catalog numbers for the standards will be:
IL1 beta human standard: ab134669
IL6 human recombinant standard: ab134672


One of my colleagues in our Cambridge UK office is adding these as we speak. It may take a few hours for the website cache to update, so please don't be alarmed if these don't show up on the website until a bit later today.

As these are new-to-us products that we have not obtained from our collaborating labs previously, there will be some backorder time from when you place the order.

I hope this is helpful. Have a fantastic weekend, and best of luck in your research!

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Answer

Thank you for your enquiry.

As the on-line datasheets indictae ab100573 is suiatbel for cell culture extracts, tissue extracts, whilst ab100572 for cell culture supernatant, urine, serum, plasma.

I would advise you totake a look at teh Protocol Booklets for these two kist which are available at our website:

https://www.abcam.com/ps/products/100/ab100572/documents/IL6-Human-ELISA-Kit-ab100572-protocol-plain.pdf

https://www.abcam.com/ps/products/100/ab100573/documents/IL6-Human-ELISA-Kit-ab100573-protocol.pdf

I hope this helps and if I can assist further, please do not hesitate to contact me.

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