製品の概要

  • 製品名

  • 検出方法

    Colorimetric
  • サンプルの種類

    Cell culture supernatant, Serum, Plasma
  • アッセイタイプ

    Sandwich (quantitative)
  • 検出感度

    < 1.5 pg/ml
  • 検出範囲

    0.66 pg/ml - 160 pg/ml
  • 添加回収試験

    96 %

    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 supernatant 97.69 85% - 104%
    Serum 96.79 84% - 103%
    Plasma 94.86 82% - 102%

  • ステップ

    Multiple steps standard assay
  • 種交差性

    交差種: Rat
  • 製品の概要

    Abcam’s IL-4 Rat ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of rat IL-4 in serum, plasma, and cell culture supernatants.

    This assay employs an antibody specific for rat IL-4 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-4 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-rat IL-4 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-4 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
    20X Wash Buffer Concentrate 1 x 25ml
    5X Assay Diluent B 1 x 15ml
    600X HRP-Streptavidin Concentrate 1 x 200µl
    Assay Diluent A 1 x 30ml
    Biotinylated anti-rat IL-4 (lyophilized) 2 vials
    IL-4 Microplate (12 x 8 wells) 1 unit
    Recombinant rat IL-4 Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • 研究分野

  • 関連性

    IL-4 is a pleiotropic cytokine produced by activated T cells. It is a ligand for interleukin 4 receptor. The interleukin 4 receptor also binds to IL13, which may contribute to many overlapping functions of this cytokine and IL13. STAT6, a signal transducer and activator of transcription, has been shown to play a central role in mediating the immune regulatory signal of IL-4. IL-4, IL3, IL5, IL13, and CSF2 form a cytokine gene cluster on chromosome 5q, with this gene particularly close to IL13. IL-4, IL13 and IL5 are found to be regulated coordinately by several long-range regulatory elements in an over 120 kilobase range on the chromosome. Two alternatively spliced transcript variants of IL-4 encoding distinct isoforms have been reported.
  • 細胞内局在

    Secreted
  • 別名

    • B cell growth factor 1
    • B cell stimulatory factor 1
    • BCGF 1
    • BCGF1
    • Binetrakin
    • BSF 1
    • BSF-1
    • BSF1
    • IL 4
    • IL4
    • Interleukin 4
    • Lymphocyte stimulatory factor 1
    • MGC79402
    • Pitrakinra
    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 ab100770 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

  • Representative standard curve using ab100770

  • Representative standard curve using ab100770

プロトコール

参考文献

ab100770 has not yet been referenced specifically in any publications.

レビューと Q&A

1-3 of 3 Abreviews or Q&A

Answer

Thank you for contacting us.

Yes, Assay Diluent A contains sodium azide which inhibits HRP activity. Therefore in this case 1X Assay Diluent B should always be used to prepare the HRP-Streptavidin and detection antibody regardless of the sample type being tested.

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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Question
Answer


The lab adviced the following:

While serum and plasma samples maybe run undiluted, we do not usually recommend doing so. The reason is not because Assay Diluent A needs to be present in every sample but that testing undiluted serum/plasma samples increases the risk of experiencing matrix effects.


Matrix effects occur when some component of the sample affects immunoreactivity of the target molecule, possibly by sequestering binding sites. Auto-antibodies, binding proteins, albumin, etc. may be possible culprits, but it’s hard to say for sure. The result is that the sample may read falsely high or low in the ELISA test, or may give a non-linear dilution response. Sometimes certain disease states contribute to matrix effects by altering the properties of the blood (highly lipemic samples, for instance); hemolysis may also cause similar problems. Matrix effects can be hard to predict and challenging to overcome.


For this reason, we rarely recommend running plasma or serum samples undiluted, as matrix effects are more likely to occur. The lowest dilution factor we normally recommend starting with is 2-fold, even for the low-abundance cytokines. If the customer is concerned that their sample levels will be below detection, keep in mind the optional overnight standard/sample incubation can be performed to try and maximize the absorbances. If that doesn’t work, the customer may want to consider concentrating their samples or switching sample types (for example plasma) to see if that helps.

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Answer


I have the following information for you regarding the kits we discussed:


ab46073:
We have no data regarding normal values observed with IL-6 and IL-4 in rat but as free circulating cytokines are rather dangerous for living animal, the normal level is probably below the detection level.


ab119548:
Unfortunately, we have not tested the rat IL-6 ELISA on normal level in rat samples. We have only tested this ELISA using samples spiked with recombinant protein.


ab100770 and ab100772:
1) What is the normal amount of rat IL4 in serum?
We tested the normal amount of IL-4 in healthy rat serum/plasma samples to be undetectable – 2 pg/ml.

2) What is the normal amount of rat IL6 in serum?
We tested the normal amount of IL-6 in healthy rat serum/plasma samples to be below the detectable limits of this ELISA. Thus, this kit may be more suitable for diseased/treated samples; however, there are some attached tips for increasing sensitivity the customer may use - see below.

3) What is the recommended dilution range for rat serum for IL4 and IL6?
We recommend a 2-fold dilution for serum/plasma samples for use with our IL-4 and IL-6 Rat ELISA kits.


Some advice for what to do if samples are reading below detection limit and how the sensitivity of the kit can be increased:


Incubate sample overnight at 4 degrees C
Increase amount of biotinylated ab (by 1.5 fold or so – too much may increase background)
Incubate biotinylated ab overnight at 4 degrees C
Increase amount of HRP-strep (by about 1.5 fold or so – too much may increase background)
Concentrate sample (using a spin column)



Please note: it’s best to try just one of these modifications at a time. Implementing too many of these changes at once may cause high background.

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