製品の概要

  • 製品名

    Mouse IL-6 ELISA Kit
    IL-6 キット 製品一覧
  • 検出方法

    Colorimetric
  • サンプルの種類

    Cell culture extracts, Tissue Extracts
  • アッセイタイプ

    Sandwich (quantitative)
  • 検出感度

    < 2 pg/ml
  • 検出範囲

    0.82 pg/ml - 600 pg/ml
  • 添加回収試験

    93 %

    特定サンプルでの回収試験
    サンプルの種類 平均 % 測定範囲
    Cell culture extracts 94.53 84% - 104%
    Tissue Extracts 93.39 83% - 103%

  • ステップ

    Multiple steps standard assay
  • 種交差性

    交差種: Mouse
  • 製品の概要

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

    This assay employs an antibody specific for mouse 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-mouse 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.

  • アプリケーション

    適用あり: Sandwich ELISAmore details
  • 試験プラットフォーム

    Microplate

製品の特性

  • 保存方法

    Store at -20°C. Please refer to protocols.
  • 内容 1 x 96 tests
    200X HRP-Streptavidin Concentrate 1 x 200µl
    20X Wash Buffer Concentrate 1 x 25ml
    2X Cell Lysis Buffer 1 x 5ml
    5X Assay Diluent 1 x 15ml
    5X Sample Diluent Buffer 1 x 10ml
    Biotinylated anti-mouse IL-6 (Lyophilized) 2 vials
    IL-6 Microplate (12 x 8 wells) 1 unit
    Recombinant mouse IL-6 Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • 研究分野

  • 機能

    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.
  • 関連疾患

    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.
  • 配列類似性

    Belongs to the IL-6 superfamily.
  • 翻訳後修飾

    N- and O-glycosylated.
  • 細胞内局在

    Secreted.
  • Information by UniProt
  • 別名

    • 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
    • Cytotoxic T cell differentiation factor
    • Hepatocyte stimulating factor
    • Hepatocyte stimulatory factor
    • HGF
    • HSF
    • Hybridoma growth factor
    • Hybridoma growth factor Interferon beta-2
    • Hybridoma plasmacytoma growth factor
    • 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
  • 参照データベース

アプリケーション

Our Abpromise guarantee covers the use of ab100713 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Sandwich ELISA Use at an assay dependent concentration.

画像

  • IL-6 measured in mouse tissue lysates prepared with RIPA buffer and sonicated in an ice-cold water bath for 12 intervals of a total of 3.5 minutes. 0.02-1 mg protein x mL-1 were tested, results shown as IL-6 per mg of extracted protein (duplicates +/- SD).

  • IL-6 measured in RIPA lysates from control cells, or cells stimulated for 24 hours with 50 ng x mL-1 of PMA (ab120297), with the addition of 1 ug x mL-1 of LPS (Sigma) (P/L) for the last 6 hours. Samples were diluted and tested in the range of 1e4-5e5 cells x mL-1, results expressed per million cells; duplicates +/- SD).

  • Dilution curves of mouse IL-6 (ab100713) and NIBSC standard (93/730). One pg of standard mouse IL-6 corresponds to 0.37 (+/- 0.07) AU. Background signal subtracted (duplicates; +/- SD).

  • Standard curve of mouse IL-6 with background signal subtracted (duplicates; +/- SD).

  • Representative standard curve using ab100713

プロトコール

参考文献

This product has been referenced in:

  • Qian X  et al. Hypouricemic and nephroprotective roles of anthocyanins in hyperuricemic mice. Food Funct 10:867-878 (2019). Read more (PubMed: 30693917) »
  • Xia YF  et al. Non-canonical Wnt signaling contributes to ventilator-induced lung injury through upregulation of WISP1 expression. Int J Mol Med 43:1217-1228 (2019). Read more (PubMed: 30664165) »
See all 22 Publications for this product

レビューと Q&A

1-6 of 6 Abreviews or Q&A

Answer

Thank you for getting in touch.

We have not tested ab100713 Mouse IL-6 ELISA Kit (Interleukin-6) specifically in liver samples in our own laboratories. However, I am happy to provide you with some general guidelines for tissue lysate preparation for this kit:

Cell or tissue lysates for this ELISA kit can be prepared using most conventional methods.  You may also use your own lysis buffer, such as RIPA or other formulations optimized for immunoprecipitation. 

Please note the following guidelines on lysis buffer composition: 

Avoid using >0.1% SDS or other strongly denaturing detergents. In general, non-ionic detergents such as Triton X-100 or NP-40 are best, although zwitterionic detergents such as CHAPS, or mild ionic detergents such as sodium deoxycholate will work. 

Use no more than 2% v/v total detergent  

Avoid the use of sodium azide 

Avoid using >10 mM reducing agents, such as dithiothreitol or mercaptoethanols 

We strongly recommend adding a protease inhibitor cocktail to the lysis buffer prior to homogenization.  Since susceptibility to proteolytic cleavage and the type of proteases present in the lysate vary, we do not recommend a specific product.  Instead, your choice of which combination of protease inhibitors to use should be based upon a literature search for your protein(s) of interest and/or tissue or cell type. Phosphatase inhibitors may be used but are not necessary unless the antibodies used in the kit specifically recognize phosphorylated forms of the protein. 


Abcam sell the following protease inhibitor cocktails:

https://www.abcam.com/products?keywords=protease+inhibitor+cocktail

Choices of the method for lysis and homogenization include glass-bead “smash,” douncing, freezethaw, sonication and crushing frozen tissue with a mortar and pestle, or even a combination of these. There is no best method for all 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, centrifuge the lysates to remove cell/tissue debris (5 min @ 10,000 x g or 10 min @ 5,000 x g) and save the supernatant. Unless testing fresh, lysates should be frozen as soon as possible and stored at -20°C (or -80°C, if possible). Centrifuge them again before incubating with anyimmunoassay. Next, determine the protein concentration of your lysates using a total protein assay not inhibited by detergents (such as the Bicinchoninic acid (BCA) assay) and normalize the volume of each sample used to deliver the same amount of total protein for each assay.  

Note: The Bradford assay is not recommended as it can be inhibited by the presence of detergents. 

Since different cells and tissues may contain different amounts of protein, as starting point, we suggest using 500 µL of lysis buffer per 1x106 cells or 10 mg tissue. You may have to adjust this based upon your results. Your target total protein concentration of the homogenate should be at least 1,000 µg/mL, but 2,000 µg/mL or more would be better.


Individual optimization of the concentration/dilution of sample in the assay may be required.

I hope this will be helpful and am happy to answer any further questions you have.

Read More

IL6 in Rat liver tissue

Good Excellent 5/5 (Ease of Use)
Abreviews
The standard curve was as described as the typical curve. One concentration had to be eliminated to give a high R^2 value.
Samples were from rat (normal out-bred sprague dawley from Harlan) liver tissue, homogenized using a 1% np-40 buffer. Although a 1:10 dilution was used for the samples (which was sufficiently in range of the IL1b ELISA (ab100768) standards), the absorbances were out of range of the standards.
For the samples that did fall in the standard range, the levels of IL6 in control rats seem to be a little higher (~225-325pg/ml) than the expected range (~125-200pg/ml).

Abcam user community

Verified customer

投稿 Apr 02 2014

Answer

Thank you for your reply.

Just to clarify, the diluted/working standards (600, 200, 66.7, 22.2, 7.4, 2.5, 0.82 pg/ml) should not be stored. The reconstituted standard (50 ng/ml) can be stored at -80ºC for 1-2 weeks at most. Remember, an extra vial of standard (Item C) is included in each kit (2 vials total) to minimize storage issues.

The example above is for theIL6 mouse ELISAkit but the same storage guidelines apply to the SDF-1a kit.
I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for your inquiry.

We know of the IL6 ELISA kit being tested in liver tissue lysate, but unfortunately we do not have any info on brain lysate.

For the SDF1 alpha ELISA kit, we haven't tested tissues with this kit but it would most likely work.If you decide to test this kit with mouse brain tissue lysates samples, we recommend diluting the samples at least 5-fold with Assay Diluent B to minimize any effects of the detergents in the lysis buffer. The samples may need to be diluted further but this would need to be determined empirically.

If you would like to test ab100741 in tissues, we do offer a testing discount program. Essentially you would purchase the kit up front at cost, test it in tissues, and submit an Abreview letting us know whether or not it works. You would then receive a discount code worth the purchase price (currently USD$470.00) off your next Abcam order within 4 months.

If you decide to test in tissues, here are some protocol recommendations.For the original lysate, in general you would want to aim for at least 1 mg/ml protein, preferably more, to achieve 50-500 ug/ml after dilution. Again though, this range should just be used as a starting point and the optimal concentration would need to be determined empirically.
In brief, a lysis buffer must meet the following specifications:
A) has relatively low salt content (700 mM or less)
B) does not contain sodium azide
C) does not contain >0.1% SDS
D) does not contain >10 mM reducing agents (beta-mercaptoethanol or dithiothreitol)
This would include any buffers used for immunoprecipitations, including RIPA buffer.
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 helps. Please contact us with any other questions.

Read More

Answer

Thank you for contacting us.

The following ELISA kits for mouse ICAM, VCAM, IL-6 and TNF-alpha are suitable for mouse samples:

ab100688 (https://www.abcam.com/ICAM1-Mouse-ELISA-Kit-1-x-96-Well-Plate-ab100688.html)
ab100713 (https://www.abcam.com/IL6-Mouse-ELISA-Kit-1-x-96-Well-Plate-ab100713.html) was tested on tissue as well.
ab100747 (https://www.abcam.com/TNF-alpha-Mouse-ELISA-Kit-1-x-96-Well-Plate-ab100747.html)
ab100750 (https://www.abcam.com/VCAM1-Mouse-ELISA-Kit-1-x-96-Well-Plate-ab100750.html)

Please let me know if these are kits you were refering to.

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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Answer

Thank you for contacting us this afternoon.

I have been to have a look at how the IL6 Mouse ELISA Kit (ab100713) looks and to take photos so that you could get a better idea of how it would work. The plate itself is a 96 well plate but can be separated into 12x8 well strips. These can be used individually when needed. I was unable to take a photo of this as the plate is packaged in a non-seethrough bag and I did not want to break the seal. I can however say that the plate is on a similar format to the following from Greiner:

http://www.greinerbioone.com/en/england/articles/catalogue/article/128_4/16891/

I hope this information has been of help. If you have any further questions please do not hesitate to contact us again.

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