Overview

  • Product name
    Human SAA ELISA Kit (SAA1)
  • Detection method
    Colorimetric
  • Sample type
    Cell culture supernatant, Serum, Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 0.5 ng/ml
  • Range
    0.41 ng/ml - 300 ng/ml
  • Recovery

    100 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 93.75 86% - 102%
    Serum 114.8 105% - 124%
    Plasma 112.1 83% - 137%

  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

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


    This assay employs an antibody specific for Human Serum Amyloid A coated on a 96-well plate. Standards and samples are pipetted into the wells and Serum Amyloid A present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human Serum Amyloid A 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 Serum Amyloid A 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. ab100635 cross reacts with SAA2.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab100635 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 ab100635.

Protocols

References

This product has been referenced in:
  • Wan-Ibrahim WI  et al. Contrasting increased levels of serum amyloid A in patients with three different bone sarcomas: An indicator of tumor malignancy? Electrophoresis 37:2328-37 (2016). Read more (PubMed: 27062367) »
  • Chang G  et al. Feeding a high-grain diet reduces the percentage of LPS clearance and enhances immune gene expression in goat liver. BMC Vet Res 11:67 (2015). Read more (PubMed: 25889631) »
See all 5 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

we have not tested this kit with urine, Please note that this does not necessarily exclude the use of urine (or any other body fluid for that matter) with this kit,

Keep in mind that our ELISA kits are very accommodating and I see no reason why urine wouldn’t be compatible so it will likely work. In general though, most protein levels are relatively lower in urine (compared to serum and plasma) so if you decide to try, then you may want to consider starting with a minimal sample dilution.








I'm sorry I'm not able to provide you with more information. Please feel free to reach out to us for any additional queries.

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Answer

Thank you for your patience whilst I talked to the lab about your results. After reviewing it, there are a few points and suggestions we can give which are summarized below

You could try a 2-fold sample dilution for your serum samples (range is 2 to 10-fold), instead of undilueted. Running serum samples undiluted can increase the risk of getting matrix effects which basically is interference by high levels of binding proteins and other serum proteins which can inhibit proper antibody-antigen binding causing inaccurately high or low signals or non-linear dilution responses. Diluting the samples can help “dilute out” these interfering molecules which can actually increase the resulting signals.

The overall standard signals are a little weak but the signal strength is still acceptable as most of the sample signals do seem to be falling within the detectable and linear range of the curve. The background and duplicate %CV are also nice and low so I would say the standard curve is fine..

The linearity of the curve is not ideal at R2 > 0.87. However, after eliminating the 300 ng/ml standard the linearity improves to R2 > 0.98.

Performing the standard/sample incubation overnight at 4ºC can boost overall signal strength so you may want to consider this next time if not done so already.

Lastly, the lower than expected concentration levels/no change between the control and the experimental samples could be the result of a couple factors. 1) The SAA levels in your are in fact just below normal. If the samples fall within the detectable range of the standard curve and the standard curve shows strong signals, low background, low duplicate %CV and high linearity than I would have high confidence that the measured values are valid. 2) Whatever treatment that is being done to the experimental samples compared to the control samples is not having as great of an impact on the SAA levels as originally thought.

I hope that helps! Please let me know if you have any questions or comments. Thanks!

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Answer

Thank you for your inquiry. Unfortunately we are unable to sell these antibodies apart from the kit. Please let me know if I can be of further assistance.

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Question
Answer

Thank you for contacting Abcam regarding ab100635.


Regarding sample volume, please use 100ul of serum. Your sample may require further dilution, but due to variations in sample, we recommend the dilution be determined by the end user.


I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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Answer

Thank you for contacting us.


Our suggested dilution fold for SAA of normal human serum is 2 – 10X. Please note that the levels of the SAA may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.


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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Answer

Thank you for contacting us.

Please find below therecommendations we obtained from the sources of the kits :

ab100635 : our suggested dilution fold for SAA of normal human serum is 2 – 10X. Please note that the levels of the SAA may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.

ab48885 : Normal human serum values obtained using the pre-coated (kit) format of ab48885 are the following :
n=40, range 1555-10 800 pg/ml, Mean 4051 pg/ml, STD 1998 pg/ml, so dilution 1:5 or 1:10 should be applied.

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

Thank you for your inquiry.  I was in contact with the lab and they do not currently have any cross-reactivity data on feline with ab100635.  If it helps though, we do know that both the capture and detection antibodies are monoclonal and that the homology between human and feline SAA is ~74% (based on online literature).  This is a bit low, as we generally say that anything with > 85% homology would be expected to cross-react. I hope this information helps. Please contact us with any other questions.

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