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.

  • 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 ab100615 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

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

  • Representative Standard Curve using ab100615.

  • 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

Customer reviews and Q&As

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1-2 of 2 Abreviews

Abreviews
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

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