Last update: 3/27/12 IK
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TP105 – Indirect 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.
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General information
Customer name:
Name of the reagent:
Catalog Number:
Lot Number:
Date received:
Storage temperature:
Was the product aliquoted?
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Specimen/sample information
Recombinant protein:
Serum:
Plasma:
Cell culture supernatant:
Other:
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Antigen coating information
Coating buffer:
Concentration:
Diluent:
Coating buffer volume/well:
Incubation time:
Incubation temperature:
Washing after coating:
Buffer:
How many times/how long:
4.
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Blocking before primary antibody information
Blocking reagent:
Volume used:
Blocking time:
Blocking temperature:
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
5.
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Primary antibody information
Catalog Number:
Lot Number:
Date received:
Storage temperature:
Was the product aliquoted?
Dilution:
Diluent:
Volume used:
Negative control for primary antibody:
Length of incubation:
Temperature of incubation:
Washing after primary antibody incubation:
Washing buffer:
How many times was washed/how long:
6.
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Secondary antibody information
Supplier:
Secondary antibody species reactivity:
Secondary antibody is conjugated with:
Dilution:
Diluent:
Negative control for secondary antibody:
Volume used:
Length of incubation:
Temperature of incubation:
Washing after secondary antibody incubation:
Washing buffer:
How many times was washed/how long:
7.
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Detection system information
Substrate:
Concentration:
Buffer:
Volume used:
Temperature:
Development time:
Absorbance wavelength:
Stopping reagent used:
8. Results
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
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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:
Positive and negative controls used on each plate:
Was the primary antibody previously working on these samples?
Was the secondary antibody previously working on other samples?
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:
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