Human Neurotrophin 3 ELISA Kit (ab100615)

Reviews (2)Q&A (3)References (1)

Overview

  • Product name

    Human Neurotrophin 3 ELISA Kit

  • Detection method

    Colorimetric

  • Sample type

    Cell culture supernatant, Serum, Plasma

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    < 4 pg/ml

  • Range

    4.12 pg/ml - 3000 pg/ml

  • Recovery

    96 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 97.25 87% - 105%
    Serum 95 85% - 103%
    Plasma 94.45 84% - 105%

  • Assay duration

    Multiple steps standard assay

  • Species reactivity

    Reacts with: Human

  • Product overview

    Abcam’s Neurotrophin 3 Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human Neurotrophin 3 in serum, plasma, and cell culture supernatants. (Human Neurotrophin 3 concentration is pretty low in normal serum/plasma, it may not be detected in this assay).


    This assay employs an antibody specific for Human Neurotrophin 3 coated on a 96-well plate. Standards and samples are pipetted into the wells and Neurotrophin 3 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human Neurotrophin 3 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 Neurotrophin 3 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    Abcam is committed to meeting high quality standards of ethical manufacturing and has decided to discontinue this product by June 2020 as it contains antibodies generated by the ascites method. An alternative kit is currently under development and will be made available as soon as possible.

  • Notes

    Optimization may be required with urine samples.

  • Platform

    Microplate

Properties

Images

  • Sandwich ELISA - Neurotrophin 3 Human ELISA kit (ab100615)
    Sandwich ELISA - Neurotrophin 3 Human ELISA kit (ab100615)

    NT-3 measured in biologicals showing quantity (pg) per mL tested sample

  • Typical Standard Curve
    Typical Standard Curve

    Representative Standard Curve using ab100615.

  • Typical Standard Curve
    Typical Standard Curve

    Representative Standard Curve using ab100615.

Protocols

References

This product has been referenced in:

  • Stewart AN  et al. Transplantation of Mesenchymal Stem Cells that Overexpress NT-3 Produce Motor Improvements without Axonal Regeneration following Complete Spinal Cord Transections in Rats. Brain Res N/A:N/A (2018). ELISA . Read more (PubMed: 29883625) »
See 1 Publication for this product

Publishing research using ab100615? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Testing Neurotrophin 3 Human ELISA Kit (AB100615) for muscle lysate samples

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
We performed 3 tests to validate the use of this ELISA kit for muscle homogenates.
In the first test we tried to use diluent A to dilute standards and samples and for tests 2 and 3 we used Diluent B. Diluent B gave us good results.

For the protocol for the sample preparation, please contact Abcam's Scientific Support Team.

Test 1. Standards and samples diluted in Diluent A
ELISA:
- Standards diluted in Diluent A.
R2 = 0.9985
Standards 1000 and 3000pg/ml were saturated. Abcam advised us to decrease incubation time for TMB One-Step Substrate Reagent to 20-25min (rather than 30min). If the values were already obtained like this, values for 3000 pg/mL (maybe even 1000 pg/mL) could be excluded based on being much higher than OD 1.0. That was done for Test 2 and 3.
- Muscle lysate samples:
Control A and B: muscles with low NT-3 expression
NT-3 A: muscle with higher NT-3 expression (injected with viral vector encoding NT-3)
- Results: See Tables 1 and 2 attached.
Poor linearity and %recovery. We then tested diluent B.

Test 2. Standards and samples diluted in Diluent B
ELISA:
- Standards diluted in Diluent B – incubation time for TMB One-Step Substrate Reagent was 25min (rather than 30min).
R2 = 0.999958
Saturation problem solved.
- Muscle lysate samples:
Control C: muscle with low NT-3 expression
NT-3 B: muscle with higher NT-3 expression (injected with viral vector encoding NT-3)
- Results:
As we homogenized samples with bigger volume for test 2 and 3, the dilution 1:2, 1:4 and 1:8 in this test is the same as 1:5, 1:10 and 1:20 from test 1.
See tables 3 and 4 attached.
NT3 samples have to be diluted further so that values can fit within the range of the standard curve. Values of the NT3 samples were lower for Diluent A probably because the recovery was too low. The % recovery could not be calculated here for NT3 samples, as it is not in the linear phase of the curve.
Control samples had very low expression and presented good % recovery. The dilution 1:8 presented worst recovery, probably because the sample is too close to the last point of the standard curve, not providing reliable results.
We decided to assay control samples diluted 1:2 and NT3 samples 1:50.
As for now, the most important for us are the NT3 samples, we did another test to validate samples diluted 1:50.

Test 3. Standards and samples diluted in Diluent B
2) ELISA
- Standards diluted in Diluent B – incubation time for TMB One-Step Substrate Reagent was 25min (rather than 30min).
R2 = 0.9996
- Muscle lysate samples:
Control D: muscle with low NT-3 expression
NT-3 C: muscle with higher NT-3 expression (injected with viral vector encoding NT-3)
- Results:
See Tables 5 and 6 attached.
Diluent B presented good linearity and good recovery for the dilution we will use for NT3 samples.
Should we do more tests? Or are the test herein enough for validation of samples diluted 1:50?

Aline Spejo

Verified customer

Submitted Aug 20 2018

Good kit

 Good Good 4/5 (Ease of Use)
Abreviews
We used the kit to detect NT3 levels in human serum. We performed the assay as per the instructions enclosed in the kit and it worked great for us.

Abcam user community

Verified customer

Submitted Jan 24 2018

Question

To whom this may concern, I would like to request for a Certificate of Compliance for Product No. ab100615 with Lot No. GR66687. Please send the certificate to this email address. Feel free to contact me with any questions or concerns regarding this email.

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Answer

Thank you for contacting Abcam. Please find attached the Certificate of Compliance for ab100615 as requested. If there is anything else I can help you with, please let me kn

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Question

I am wondering whether the NT-3 ELISA kit you offer (ab100615) is compatible with cell lysate lysed with the following buffer components: 20mM Tris-HCl pH7.4, 1mM EDTA, 1mM EGTA, 1% NP40, PI?

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Answer

 Thank you for your inquiry. We haven't tested ab100615 with cell lysates.  It would most likely work, but we wouldn't be able to guarantee it since it is untested.   If you decide to test this kit with lysate samples, we recommend diluting the samples at least 5-fold with Assay Diluent B to minimize any effects of the detergents in the lysis buffer.  The samples may need to be diluted further but this would need to be determined. The lysis buffer should: A) have relatively low salt content (700 mM or less) B) not contain sodium azide C) not contain >0.1% SDS D) not contain >10 mM reducing agents (beta-mercaptoethanol or dithiothreitol)   This would include any buffers used for immunoprecipitations, including RIPA buffer.   I hope this information helps.  Please contact us with any other questions.      

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Question

Does ab100615 cross-react with mouse? I would like to test ab75960 and ab28976 as a sandwich ELISA pair.

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Answer

Thank you very much for your interest in ab100615. This kit has not been validated for use with mouse samples, but is predicted to react based on high sequence homology. I can offer a discount off a future purchase if you buy either this ELISA kit or pair of antibodies now, test in sELISA on mouse samples and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free kit for testing the ELISA kit, or two free antibodies for testing the sandwich ELISA pair. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know which item(s) you would like to test. I will then send a discount code. This code must be issued before purchasing the products so please wait for my reply before ordering. 2. Purchase ab28976 and ab75960 or ab100615 either by phone, fax, or online (www.abcam.com). 3. Test it in sELISA on mouse samples. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the product turns out to be unsuitable for sELISA on mouse samples, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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