Last update: 3/27/12 IK
Document1
Page 1 of 3
TP106 – Sandwich ELISA Troubleshooting
In order for us to help you better and faster, please provide us with as much information as
possible by filling out this form and returning to Abbiotec Technical Support Department by
email to techsupport@abbiotec.com or fax to 1 (858) 586-6252. Attach any relevant
experimental data (positive or negative) with legend.
1.







General information
Customer name:
Name of the reagent:
Catalog Number:
Lot Number:
Date received:
Storage temperature:
Was the product aliquoted?
2.





Specimen/sample information
Recombinant protein:
Serum:
Plasma:
Cell culture supernatant:
Other:
3.



Standard information
Reconstitution volume of standard
Reconstitution medium
Reconstitution time (before adding standard to the wells)
4.







Capture antibody information
Catalog Number:
Lot Number:
Date received:
Storage temperature:
Was the product aliquoted?
Capture antibody is conjugated with:
Dilution:
Abbiotec, LLC • 7985 Dunbrook Rd, Ste A, San Diego, CA 92126 • USA
Ph: 858.586.0500 • Fax: 858.586.6252 • Email: info@abbiotec.com • WWW.ABBIOTEC.COM
Last update: 3/27/12 IK
Document1
Page 2 of 3








Diluent:
Negative control for capture antibody:
Volume used:
Length of incubation:
Temperature of incubation:
Washing after capture antibody incubation:
Washing buffer:
How many times was washed/how long:
5.










Capture antibody coating information
Coating buffer:
Concentration:
Diluent:
Coating buffer volume/well:
Incubation time:
Incubation temperature:
Washing after coating:
Buffer:
Volume used:
How many times/how long:
6.







Blocking after capture antibody information
Blocking reagent:
Blocking time:
Blocking temperature:
Washing after blocking:
Buffer:
Volume used:
How many times/how long:
7.










Detection antibody information
Catalog Number:
Lot Number:
Date received:
Storage temperature:
Was the product aliquoted?
Detection antibody is conjugated with:
Dilution:
Diluent:
Negative control for detection antibody:
Volume used:
Abbiotec, LLC • 7985 Dunbrook Rd, Ste A, San Diego, CA 92126 • USA
Ph: 858.586.0500 • Fax: 858.586.6252 • Email: info@abbiotec.com • WWW.ABBIOTEC.COM
Last update: 3/27/12 IK
Document1
Page 3 of 3





Length of incubation:
Temperature of incubation:
Washing after detection antibody incubation:
Washing buffer:
How many times was washed/how long:
8.








Detection system information
Substrate:
Concentration:
Buffer:
Volume used:
Temperature:
Development time:
Absorbance wavelength:
Stopping reagent used:
9.





















Results
Please describe your results briefly:
Is there a color reaction?
No signal or weak signal:
Are ODs normal or too low?
High background:
Uneven color development:
Poor standard curve:
Positive and negative controls used on each plate:
Antibodies only control (no standards):
Reagents only control (no antibodies, no standards):
Fresh buffers and substrates were used:
Reagents and buffers were stored at 4°C and used at working temperature:
Was the right microtiter plate use?
Microtiter plates covered during incubations:
Was the capture antibody previously working on these samples?
Was the detection antibody previously working on other samples?
Washing steps were done correctly:
ELISA reader is functioning well, correct wavelength was used for reading:
How many times this reaction was replicated?
Are the results the same every time?
Please send us a copy of raw data for each plate.
Abbiotec, LLC • 7985 Dunbrook Rd, Ste A, San Diego, CA 92126 • USA
Ph: 858.586.0500 • Fax: 858.586.6252 • Email: info@abbiotec.com • WWW.ABBIOTEC.COM