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    rat-il-4-elisa-kit-ab100770.pdf

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Immunology Adaptive Immunity T Cells Non-CD
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Rat IL-4 ELISA Kit (ab100770)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (3)References (2)

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Typical standard curve
  • Typical standard curve

Key features and details

  • Sensitivity: 1.5 pg/ml
  • Range: 0.66 pg/ml - 160 pg/ml
  • Sample type: Cell culture supernatant, Plasma, Serum
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Rat

You may also be interested in

Protein
Recombinant rat IL-4 protein (ab9812)

View more associated products

Overview

  • Product name

    Rat IL-4 ELISA Kit
    See all IL-4 kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 1.5 pg/ml
  • Range

    0.66 pg/ml - 160 pg/ml
  • Recovery

    96 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 97.69 85% - 104%
    Serum 96.79 84% - 103%
    Plasma 94.86 82% - 102%
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Rat
  • Product overview

    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.

  • 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
    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
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • Non-CD
    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cytokines and cytokine receptors ELISA kits
  • Relevance

    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.
  • Cellular localization

    Secreted
  • Alternative names

    • 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
  • Database links

    • Entrez Gene: 287287 Rat
    • SwissProt: P20096 Rat

    Associated products

      Images

      • Typical standard curve
        Typical standard curve

        Representative standard curve using ab100770

      • Typical standard curve
        Typical standard curve

        Representative standard curve using ab100770

      Protocols

      • Protocol Booklet

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet
      • SDS
    • References (2)

      Publishing research using ab100770? Please let us know so that we can cite the reference in this datasheet.

      ab100770 has been referenced in 2 publications.

      • Eftekhar N  et al. Immunomodulatory and anti-inflammatory effects of hydro-ethanolic extract of Ocimum basilicum leaves and its effect on lung pathological changes in an ovalbumin-induced rat model of asthma. BMC Complement Altern Med 19:349 (2019). PubMed: 31801507
      • Achi SC  et al. Prophylactic effects of probiotic Bifidobacterium spp. in the resolution of inflammation in arthritic rats. Appl Microbiol Biotechnol 103:6287-6296 (2019). PubMed: 31168650

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-3 of 3 Abreviews or Q&A

      Question

      Can serum samples be run "neat" (without dilution) in the IL-4 Elisa - ab100770?

      Read More

      Abcam community

      Verified customer

      Asked on May 16 2013

      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.

      Read More

      Abcam Scientific Support

      Answered on May 16 2013

      Question

      regarding IL-4 rat elisa (ab100770) -

      In the protocol (page 7) #6. PREPARATION OF REAGENTS, step 6. says to add diluent B to prepare the detection antibody and step 7 says to add diluent B to dilute HRP. My question is this, should you use diluent B regardless of whether your samples are serum/plasma (diluted with diluent A) or cell culture medium (diluted with diluent B)?



      Thank you

      Read More

      Abcam community

      Verified customer

      Asked on Jun 03 2013

      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.

      Read More

      Abcam Scientific Support

      Answered on Jun 03 2013

      Question

      What is the normal amount of rat IL4 in serum?
      What is the normal amount of rat IL6 in serum?
      What is the recommended dilution range?

      Read More

      Abcam community

      Verified customer

      Asked on May 13 2013

      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.

      Read More

      Abcam Scientific Support

      Answered on May 13 2013

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