About Elisa Troubleshooting Tips
About Elisa Troubleshooting Tips
The Elisa Troubleshooting Tips is somewhat seeking to experience the fruitful outcome. Let’s
improve Troubleshooting with the help of the following tips:
1. In a case low standard curve
To enhance standard solution, try to confirm that dilutions are made accurately, if not,
kindly amend it.
If curves are not fitting up to the required scale, make sure to try plotting using various
scales such as 5 parameters logistic curve fit.
In a case when standard improperly reconstituted, then slowly spin vial before opening
as well as examine for not dissolved material after reconstituting.
In a case of pipetting error, if so, then, use calibrated pipettes and proper pipetting
2. In the case of No Signal
If Incubation time too small, then incubate samples overnight at 4°C.
If incompatible sample type, then make sure to reduce the deductions. You may include a
sample that will help you to find out a positive control.
If target presents under detection limits, then decline dilution factor.
If samples, prepare inaccurately, then make sure a proper sample dilution. Moreover,
samples may be unsuited with microtiter plate assay format.
In a case of not enough detection reagent, then hike up the amount of detection reagent
If reduced antibody, then try various dilutions of antibody.
If incubation temperature is too less, then make sure incubations are run at the
accurate temperature. Moreover, in this, all the reagents should be at room
If inaccurate wavelength, then first examine the wavelength.
If the plate is washing too rough, then have a look at the pressure point to set it
accordingly. Moreover, pipette washes buffer softly if washes are done manually.
If the slow color development of enzymatic reactions, then make sure to prepare a
substrate solution immediately before use. Moreover, keep checking that the stock
solution has not expired.
3. In case of the Large coefficient of variation
To enhance the overall performance, make sure to follow this Sandwich Elisa Troubleshooting
If find about the bubbles in wells, then make sure no bubbles are present before to
If incomplete reagent mixing, then make sure that all the reagents are mixed
If inconsistent pipetting, then indulge in calibrated pipettes and systematic technique to
provide reliable pipetting.
If inconsistent sample preparation, then provide consistent sample preparation and
maximum sample storage conditions.
In the case of High Background
If massive detection reagent, then provide the reagent has been diluted entirely.
If the contaminated wash buffer, then arranges freshwater buffer.
If high backgrounds are not properly washed, then make sure to wash well as per
4. In the case of Low sensitivity
If insufficient target, then concentrate on reducing sample dilution.
If improper storage of ELISA kit, then make sure to keep all reagents as recommended.
If target badly adsorbs to microtiter, then covalently link target to microtiter.
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