Anti-IGF1 antibody (ab9572)

To our knowledge, this IGF antibody has not been tested in flow cytometry and our guarantee does not cover this application of the antibody. However it has been tested and shown to work in a similar application, immunocytochemistry, after methanol fixation of the cells. For flow cytometry, your cells will need to be fixed and permeabilized, as IGF is completely intracellular until it is secreted.

Thank you for contacting us.

The lab sent me the following information:

The immunizing antigen was produced in E.coli.
The purification of this antibody is done using an antigen affinity column.
This is all the information we have available.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Thank you for your message and for providing this further information.

To confirm, I can confirm this antibody has been tested and therefore guaranteed for human samples. The previous email stated that it has not been tested specifically with serum samples in ELISA. Serum contains many other proteins and other biological factors that may be interfering with the antibody binding.

I am sorry to hear the suggestions made have not improved the results on this occasion. As previously discussed,the antibodies seem to be workingbut regrettably there seems to be some interference from something within the serum samples. I appreciate the time you have spent on these experiments and as I gesture of goodwill I would be pleased to arrange a credit note. Alternatively, Iwould be pleased to provide free of charge replacements if this is what you prefer and would like to try again. However, please bear in mind it is possible this may not solve the issue if there is a possibility the serum sample type is not suitable for use with these antibodies in ELISA.

I hope this will provide a satisfactory resolution. I look forward to hearing from you with details of how you would like to proceed.

Thank you for your reply

Please note that theseantibodies have not been tested with with serum samples, and so I am sorry no pretreatments have been tested which we could provide you with information on. In theory the antibodiesshould work with serum samples. Since the antibodies are working well with their standard protein, this means that the antibodies do work in the ELISA.Therefore, I can suggestit may be that there is some interferencewith something in the serum samples.

1. Have you been able to confirm the expected amount of protein in their samples using any other method?

2. Is it possible that the samples contain a very small amount of protein?

3. Is there any azide in the samples? This could inhibit the reactionand would lead to loss ofdetection as you are observing.

4. Have you tried any other sample types?

5. Please provide details of the sample preparation.

I hope this will be helpful. Please do not hesitate to get back to me with the requested information and I hope we can resolve this for you as soon as possible.

Thank you for your reply.

I would like to reassure you that ab83137 Anti-IGF1 antibody (Biotin) and ab9572 Anti-IGF1 antibody are tested and covered by our 6 month guarantee for sandwich ELISA together, and human samples. 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.

As the difficulties are continuing, I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you could include details of the sample pretreatment. What was the pretreatment protocol, and the reasons why this was done?

I would appreciate if you could also providethe data from the standard curve and samples which would help us to assess the results.

With regards to the protocol used for testing, both the IGF1 and IGFBP3 antibodies will have been tested using the same procedure. Once I have further information from you, I would like to investigate further this further with our source if they have any other recommendations for you from their experiences with this antibody, particularly with regards to sample type and preparation.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.


Order Details
Antibody code:

Problem
Choose: Problem with standard curve No signal or weak signal High background


Lot number


Purchase order number
or preferably Abcam order number


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, no signal, non-specific color development, poor standard curve etc.)


Type of ELISA (Direct ELISA/Indirect ELISA/Sandwich ELISA etc.)


Sample (Species/Cell type/Cell line etc.)


Coating well (Buffer/Concentration of the coating material etc.)


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 (Substrate/Diluent etc.)


Positive and negative controls used (please specify)




Optimization attempts (problem solving)
How many times have you tried the ELISA?



Have you run a "No Primary" control?


Yes No
Do you obtain the same results every time?
Yes No

What steps have you altered?


Additional Notes:



Data:
We would appreciate if you are able to provide a copy of the data, particularly from the standard curve. This will help us to assess the results.