Overview

  • Product name

    Human IL-15 ELISA Kit
    See all IL-15 kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 3 pg/ml
  • Range

    3.3 pg/ml - 800 pg/ml
  • Recovery

    94 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 96.37 83% - 104%
    Serum 92.26 80% - 102%
    Plasma 94.53 82% - 103%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s IL-15 Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human IL-15 in serum, plasma and cell culture supernatants.


    This assay employs an antibody specific for Human IL-15 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-15 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human IL-15 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-15 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    Get results in 90 minutes with Human IL-15 ELISA Kit (ab176110) from our SimpleStep ELISA® range.


     

  • Notes

    Optimization may be required with urine samples

  • 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
    200X HRP-Streptavidin Concentrate 1 x 200µl
    20X Wash Buffer Concentrate 1 x 25ml
    5X Assay Diluent B 1 x 15ml
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Human IL-15 (lyophilized) 2 vials
    IL-15 Microplate (12 x 8 wells) 1 unit
    Recombinant Human IL-15 Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

  • Relevance

    IL-15 (114 amino acids) is a cytokine that regulates T and natural killer cell activation and proliferation. It has a predicted molecular mass of approximately 12.5 kDa. Human IL-15 shares approximately 97% and 73% amino acid sequence identity with simian and mouse IL-15, respectively. Both human and simian IL-15 are active on mouse cells. IL-15 was initially isolated from the simian kidney epithelial cell line CV1/EBNA. It has also been isolated from mouse and human cell sources. The cytokines IL-15 and IL2 share many biological properties and stimulatory activities (T, B, and NK cells). Both IL-15 and IL-2 stimulate mouse CTLL2 cells. In activated peripheral blood T lymphocytes, IL-2 is highly expressed but the expression of IL-15 is not detectable. There is no sequence homology between IL-15 and IL-2, though computer modeling indicates both possess a four alpha helical bundle structure. IL-15 competes for binding sites with IL-2, as both IL2 and IL-15 stimulate the growth of cells through the IL-2 receptor. IL-15 mRNA is expressed in many cell types and tissues including adherent peripheral blood mononuclear cells, fibroblasts, and epithelial cells, monocytes, placenta, and skeletal muscle.
  • Cellular localization

    Isoform IL-15-S48AA: Secreted. Isoform IL-15-S21AA: Cytoplasm. Nucleus. Note: IL-15-S21AA is not secreted, but rather is stored intracellularly, appearing in the nucleus and cytoplasmic components.
  • Alternative names

    • IL 15
    • Interleukin 15
    • Interleukin15
    • MGC9721
  • Database links

Applications

Our Abpromise guarantee covers the use of ab100554 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 ab100554

  • Representative standard curve using ab100554

Protocols

References

This product has been referenced in:

  • Yamamoto M  et al. Stage classification of IgG4-related dacryoadenitis and sialadenitis by the serum cytokine environment. Mod Rheumatol N/A:1-5 (2018). Read more (PubMed: 29385874) »
  • Iki H  et al. Effect of exercise therapy on cytokine secretion in the saliva of bedridden patients. J Phys Ther Sci 28:2871-2876 (2016). Human . Read more (PubMed: 27821953) »
See all 2 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

easy IL15 ELISA

Good Excellent 5/5 (Ease of Use)
Abreviews
IL15 was measured in human culture supernatants, diluted with assay diluent B as recommended.
Solid standard curve and very easy to use kit. However, according our ODs of the standards, the kit is not quite as sensitive as stated.

Dr. Anna Vossenkaemper

Verified customer

Submitted Jun 30 2015

Answer

’ai contacté le laboratoire mais malheureusement comme expliqué par téléphone, nous n’avons pas testé ce kit avec Il-15 attaché au récepteur.

De plus, puisque nous ne connaissons pas l’épitope des anticorps utilisé danse ce kit, il est difficile de savoir si l’interaction Il-15-recepteur affectera les résultats.

Read More

Answer

I am getting back to you in regards to the problems experienced when using this kit.

We have not yet encountered an issue with our ELISA kits that were caused by not printing the capture antibodies. All of our kits are pre-coated.

It is very unlikely that the signal produced from the samples comes from non-specific binding alone. This would imply the background response from the samples is extremely high, some above 1.5 OD. Because of the wash steps involved in the procedure, these readings are simply too high to be considered only non-specific binding to the plastic wells.

According to my lab colleagues there may have been an issue with the standard preparation, or the standard itself had degraded.

If possible, would it be possible to test the second, fresh vial of standard recombinant protein provided with the tips provided?

I have requested the standard OD values we obtained from testing this lot. I will send them to you as soon as I receive them.

Please let me know if you need further assistance.

Read More

Answer

Thank you for contacting us.

I appreciate your patience and apologize for keeping you waiting.

As none of the standards gave strong signal responses it could be that the standard recombinant protein has degraded. However, because the results obtained with the R&Ds recombinant human protein were identical, it may suggest an issue with the standard preparation.

In my opinion the pre-coated microplate could not have caused these results, because the samples gave good, strong signals.

I would encourage you to use the second, fresh vial of Standard Recombinant IL-15 keeping in mind the following tips for ensuring strong signals:

When preparing your standards, it is very critical to briefly spin down the vial first. The powder may drop off from the cap when opening it if you do not spin down. Be sure to dissolve the powder thoroughly when reconstituting. After adding Assay Diluent to the vial, we recommend inverting the tube a few times, then flick the tube a few times, and then spin it down; repeat this procedure 3-4 times. This is a technique we find very effective for thoroughly mixing the standard without too much mechanical force.
Do not vortex the standard during reconstitution, as this will destabilize the protein.
Once your standard has been reconstituted, it should be used right away or else frozen for later use.
Keep the standard dilutions on ice while during preparation, but the ELISA procedure should be done at room temperature.
Be sure to discard the working standard dilutions after use – they do not store well.



I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you for contacting us.
I am very sorry to hear this kit failed to give the expected results. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
In order to better understand the problem, I’d like to gather further information. I would really appreciate if you could please clarify the following:
- How was the kit stored? And how was the standard stored after reconstitution?
- You mention this kit was successfully used in the past. Could you please provide the lot number which worked fine previously?
- Which assay diluent was used (A or B) with your standard / samples?
I am happy to investigate this case for you, and the event that the kit sent happened to be faulty, I will replace o reimburse it as stated by the Abpromise guarantee.
I look forward to receiving your reply. Have a nice day.

Read More

Answer

Thank you for contacting us.

Since the protocol recommends to store the concentrateat 4°C for 5 days, this is what would be best. If the concentrate is still good after 21 days and how good it is, that would need to be tried out and is difficult to predict.

On the other hand if you want to be on the safe sideand your samples are very precious, I would suggest using the second vial of item F instead.

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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