Kit Performance Information
Protein Carbonyl Assay Kit Performance
Our technical support group has provided the following information on the performance of the kit.
nmol/mg standard | CV | Back-calculated concentration | Deviation |
|---|---|---|---|
0.32 | 5.7% | 0.31 | -4.9% |
0.65 | 3.1% | 0.65 | 0.0% |
1.07 | 3.6% | 1.01 | -5.0% |
1.31 | 4.6% | 1.37 | 5.2% |
1.72 | 5.5% | 1.71 | -0.9% |
Intra-assay precision and accuracy
6 wells for each sample
Inter-assay precision
Quality control sample | Low | Medium | High |
|---|---|---|---|
Number of assays | 6 | 6 | 6 |
Mean (nmol/mg) | 0.23 | 0.65 | 1.26 |
SD | 0.03 | 0.03 | 0.08 |
CV | 12.2 | 4.5 | 6.1 |
nmol/mg standard
0.05
0.1
0.25
0.5
1.0
CV
4.0%
3.6%
5.0%
1.7%
2.8%
0.068
0.114
0.243
0.507
0.996
back-calculated
concentration
+36%
+14%
-2.8%
+1.4%
-0.4%
CV
nmol/mg
0.05
0.25
0.5
1.0
CV
3.6%
3.7%
2.9%
4.2%
nmol/mg result range
<0.1
0.11-0.3
>0.3
Number of samples
18
9
12
4.6
2.9
3.8
Average CV%
Quality Control
Sample
Number of Assays
Mean
SD
CV
Medium
6
0.201
0.016
7.7%
6
0.560
0.033
5.8%
High
6
0.050
0.012
24.7%
Low
This selection of samples lies within the range of levels commonly found in clinical samples, e.g. plasma. In general, at the low end of the standard curve, a higher variation is expected which decreases as levels increase.
Intra-assay variation for clinical samples (plasma)
3 wells per sample
nmol/mg result range | Number of samples | Average CV% |
|---|---|---|
0.16-0.26 | 26 | 8.41% |
Biotin hydrazide (new) vs Dinitrophenylhydrazine (original) assay:
The original protein carbonyl assay (formerly marketed by BioCell) was based upon derivatisation of protein carbonyls with dinitrophenylhydrazine (DNP), followed by probing with an anti-DNP-biotin antibody, and then subsequent detection with streptavidin-biotinylated horseradish peroxidase.
The new version of the assay is based on the same principle but uses biotin hydrazide as the derivatisation agent, followed by detection with streptavidin-biotinylated horseradish peroxidase. This antibody-free approach results in a simpler assay that is quicker to perform and has reduced non-specific background noise.
A selection of plasma samples (n = 26) were compared across the new and original assays. While samples exhibited similar trends across both assays, samples measured using the new biotin hydrazide-based assay were at or above the limit of quantification (LOQ), whereas samples measured in the DNP-based assay were around the limit of detection (LOD) and well below the limit of quantification. The LOD and LOQ values are similar for both the biotin hydrazide-based assay (shown on graph) and the DNP-based assay (not shown).