Sample
Serum, plasma, tissue homogenates
Intended Use
For the quantitative determination of human receptor tyrosine-protein kinase erbB-3 (ERBB3) concentrations in serum, plasma, tissue homogenates.
Contents of Kit
1. Assay plate (12 × 8 coated Microwells), 1(96 wells)
2. Standard (Freeze dried), 2
3. Biotin-antibody (100× concentrate), 1 × 120 μl
4. HRP-avidin (100× concentrate), 1 × 120 μl
5. Biotin-antibody Diluent, 1 × 15 ml
6. HRP-avidin Diluent, 1 × 15 ml
7. Sample Diluent, 1 × 50 ml
8. Wash Buffer (25× concentrate), 1 × 20 ml
9. TMB Substrate, 1 × 10 ml
10. Stop Solution, 1 × 10 ml
11. Adhesive Strip (For 96 wells), 4
12. Instruction manual, 1
Storage
Unopened kit: Store at 2 - 8°C. Do not use the kit beyond the expiration date.
Opened kit:
Standard/Biotin-antibody/HRP-avidin: May be stored for up to 1 month at 2 - 8° C. If don' t make recent use, better keep it store at -20°C.
Biotin-antibody Diluent/HRP-avidin Diluent/Sample Diluent/Wash Buffer/TMB Substrate/Stop Solution: May be stored for up to 1 month at 2 - 8°C.
*Provided this is within the expiration date of the kit.
Precision
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Detection Range
125 pg/ml-8000 pg/ml
Sensitivity
The minimum detectable dose of human ERBB3 is typically less than 31.25 pg/ml.
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined the mean O.D value of 20 replicates of the zero standard added by their three standard deviations.
Standard Curve
Using the professional soft "Curve Expert" to make a standard curve is recommended, which can be downloaded from our web.
Average the duplicate readings for each standard and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the ERBB3 concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data.
If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
Citations
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