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Dear Sir/ Madam,
We are trying to develop and ELISA for Dengue NS1 detection and having been working with your products in some of our previous and other present work, we have considered using your dengue products as well. However, during our preliminary experiments, we have noticed that the dengue NS1 antigen (ab64456) cannot be detected by either the anti-dengue NS1 antibody (ab41616) that you are also suggesting for other costumers in the technical support page, or the anti-histidine tag antibody (ab9136). This has led us to decide for a replacement of the antigen NS1 antigen (ab64456).
Moving forward, we outsourced another dengue NS1 antigen from a different supplier and also an anti-dengue NS1 antibody which has been used in ELISA and WB applications for the detection of native NS1 antigens. (The antibody reacts with the 3rd and 4th conformational epitope of the Dengue NS1 based on the immunogen preparation.) We further tested the anti-dengue NS1 antibody (ab41616) using the new NS1 antigen and comparing with the new anti-dengue NS1 antibody as well. However, anti-dengue NS1 antibody (ab41616) still does not react with the new dengue NS1 antigen even at higher concentrations of 1:250. In comparison, the new antibody detected the dengue NS1 antigen adsorbed, and is also detected and validated by the anti-histidine tag antibody (ab9136).
Since, it took a little time for us to outsource another dengue NS1 antigen to further test the reactivity of the anti-dengue NS1 antibody (ab41616) and perform evaluation, we are very much sorry for the delay but we would like to request for the possibility of replacement for the anti-dengue NS1 antibody (ab41616) into Anti-E.coli antibody (ab25823) that we would like to use for our other project.
We are looking forward for a positive response from you. Thank you in advance.
Asked on Jul 13 2012
Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.
I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.
I would like to reassure you that this antibody is tested and covered by our guarantee for WB. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality records to determine what may be the cause of the problem.
In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I have completed the sections for which information has already been provided.
It would also be beneficial if you are able toconfirm:
1. What other samples have been tested with this antibody? Has it detected endogenous protein in any samples? I can suggest it would be beneficial to try this if possible.
2. Has the protein been stained with comassie on an SDS gel? Is it intact and running at the correct molecular weight?
In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.
Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.
Choose: Non-specific band Multiple bands No signal or weak signal High background
Purchase order number
or preferably Abcam order number:
Antibody storage conditions (temperature/reconstitution etc)
Description of the problem (high background, wrong band size, more bands, no band etc.)
Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Amount of protein loaded
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Detection method (ECL, ECLPlus etc.)
Positive and negative controls used (please specify)
Optimization attempts (problem solving)
How many times have you tried the Western?
Have you run a "No Primary" control?
Do you obtain the same results every time?
e.g. are the background bands always in the same place?
What steps have you altered?
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.
Answered on Jul 13 2012