Overview

  • Product name

    Human Fas Ligand ELISA Kit (APTL)
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 2 pg/ml
  • Range

    1.37 pg/ml - 1000 pg/ml
  • Recovery

    75 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 76.99 70% - 90%
    Serum 76.36 70% - 87%
    Plasma 73.42 67% - 86%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

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

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

  • Notes

    Optimization may be required with urine samples

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab100515 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 ab100515.

  • Representative Standard Curve using ab100515.

Protocols

References

This product has been referenced in:

  • Zhang Z  et al. Astragalus mongholicus (Fisch.) Bge Improves Peripheral Treg Cell Immunity Imbalance in the Children With Viral Myocarditis by Reducing the Levels of miR-146b and miR-155. Front Pediatr 6:139 (2018). Read more (PubMed: 29977885) »
See 1 Publication for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

good

Good Excellent 5/5 (Ease of Use)
Abreviews
This is a good Elisa.
The results are good and reproducible.
The protocol is well explained and easy to understand and easy to implement.
We used Serum samples.

Abcam user community

Verified customer

Submitted Apr 17 2018

good working Elisa

Good Excellent 5/5 (Ease of Use)
Abreviews
This is a good Elisa.
The results are good and reproducible.
The protocol booklet is well explained and easy to understand and easy to perform.
I have only used 1 kit for measuring Fas-Ligand so far.
I used this kit for serum samples from a comparison of healthy donor and atopic patients.
The kit is easy to use and quick compared to other Sandwich Elisa.
Based on the recommendations within the booklet we used a serum dilutions of 1: 100 and these dilutions was within the linear range for all but of the 40 samples.

Abcam user community

Verified customer

Submitted Apr 08 2015

Question
Answer

We would suggest testing a range of dilutions to find a dilution within the standard curve. A good range is neat, 1:5, 1:10, 1:20. In the protocol the suggested dilution for normal serum/plasma: 2-10 fold, again this needs to be optimized.

Read More

Answer

Unfortunately, the OD standard is never twice the same, therefore, it is so important to run a fresh standard curve for EVERY assay. The OD needs to be measured, it cannot be not calculated.
How to analyze the data is described under section 8, page 11 of our protocol booklet (see attachment).
As we haven't tested saliva samples, we cannot make a proper suggestion. I would suggest to test this yourself.

According to the protocol booklet, the storage and handling is described in section 4:
ab100515 may be stored for up to 6 months at 2 to 8°C from the date of shipment. Standard (recombinant protein) should be stored at -20°C or -80°C (recommended at -80°C) after reconstitution.
Opened Microplate Wells or reagents may be store for up to 1 month at 2 to 8°C. Return unused wells to the pouch containing desiccant pack, reseal along entire edge. Note: the kit can be used within one
year if the whole kit is stored at -20°C. Avoid repeated freeze-thaw cycles

Read More

Answer

The exact epitopes are not known but the antibody pair is binding somewhere on the extracellular domain (aa 134-281). If it helps, the capture antibody is monoclonal and the detection antibody is polyclonal. We do not have a kit against HA although we have many antibodies that are anti-HA.

Read More

Answer

I can confirm that these kits will detect both soluble and membrane bound Fas and fas ligand.

In order to prevent membrane bound protein to interfere with your results, I suggest to spin down the saliva samples to get rid of all cells while taking care not to generate cell debris.

Plasma is is in generally cell free and therefore should not contain any membrane bound proteins.

Read More

Answer

I have reviewed the details on the datasheets for these kits. For the Fas ligand kits:

ab45892 Fas Ligand Human ELISA Kit
ab100515 Fas Ligand Human ELISA Kit

Sensitivities are different:ab100515 < 2 pg/ml, ab45892 < 12 pg/ml

Detection ranges are different:ab10515 1.37 pg/ml - 1000 pg/ml,ab45892 62.5 pg/ml - 2000 pg/ml

For the CD95 kits:
ab45922 CD95 Human ELISA Kit
ab100513 CD95 Human ELISA Kit

Again, sensitivities are different: ab100513 < 5 pg/ml, ab45922 < 47 pg/ml
Detection ranges are different:ab45922 93.75 pg/ml - 3000 pg/ml, ab100513 2.74 pg/ml - 2000 pg/ml

These are the main differences you may need to take into consideration. However, I can recommend to review the datasheets fully for further information and otherdifferences.

Read More

Answer

These ELISA kits are from two different sources but they are similar.

I recommend to determine which is the best for your needs from the datasheet and protocol information. Please take not of the different ranges these kits are suitable for. I suggest to choose the kits that are most suitable for the amounts of Fas ligand and CD95 you are expecting in your samples.

I believe that all these kits are suitable for un-bound (free) Fas ligand and CD95 since for ELISA the cells are lysed and ELISA is not a cell based assay.

Read More

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