Anti-CD3 antibody (ab5690)

Thank you for taking the time to complete our questionnaire and send the images. The details you have kindly provided will provide us with vital information for our monitoring of product quality

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there are not many further tips to provide on this occasion to help improve the results. I can suggest you have received a bad vial on this occasion.

I apologize for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation, providing the item has been purchased in the last 6 months and the antibody has been usedin a tested species. In order to arrange this for you, I would appreciate if you could confirm:

1. The order number and date of purchase.

2. The species which has been tested.

In addition, I can suggest you may like to consider the following suggestions as a check for future experiments:

1. This antibody not previously tested with methacarn fixation before. It may be that it is not suitable for this antibody. The Abreviews for IHC on frozen sections submitted by other customers on the datasheet have used PFA fixation or acetone. I can suggest to consider trying these.

2. Is the secondary antibody working well with other primary antibodies? What are the results of the no primary control?

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed and with the requested information.

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 IHC-P, IHC-Fr and for mouse, rat and human samples. 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.

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.

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 and hope we can resolve this for you as soon as possible.

Order Details

Antibody code:

Lot number:

Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific satining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)


Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


Antigen retrieval (Enzymatic method, Heat mediated technique etc.)


Permeabilization step


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


Positive and negative controls used (please specify)


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



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


We would appreciate if you are also able to provide and image which woudl help us to assess the results

Thank you for contacting us.



I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Thanks for your enquiry. I am sorry this vial of ab5690 is giving you trouble.

Please feel free to contact me when you have decided if you would like a replacement, credit or refund. As a reminder, please include your PO number or Abcam order reference number in your reply so I can locate your initial order and process your request.

Thanks!

Thank you very much for sending all of this information.

I'm sending free of charge vials of ab39626 and ab39628 on the order ***, which should arrive on Monday.

Just to confirm, the macrophages were from mice or a different species? We haven't tested ab119774 with mouse samples, so wecan't guarantee that it reacts with the mouse protein. If you'd ever like to try an antibody with an untested species, we do have a testing discount program that you might be interested in. Feel free to contact me if you'd ever like more information.

If you have any questions or if there is anything else that we can do for you, please let me know and I'll be happy to help. Have a nice weekend.

Thank you so much for contacting us and for letting us know about the trouble with these two antibodies.

We do fully guarantee our products to work as stated on the datasheets in tested species and applications, so if the antibodies did not work as they should have, we can provide replacements. For our records, could you please tell me the following information?

1) What application were the antibodies used in? What kind of results were seen (high background, no staining/bands, etc)?

2) What species were the samples from? What kind of tissue/cells were used?


3) Could you describe the protocol used? Please include the antibody dilution/concentration and components of the blocking solution.

I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you, and I'll be happy to help.

Thanks for your enquiry.

Here are some tips that I hope will reduce the background in your IHC:

1. Increase the dilution of primary antibody from 1:100to 1:300

2. Increase the dilution of secondary from 1:500 to 1:1000

3. Consider using a "no primary" control experiement to deterimine if the background is due to the secondary

4. Use the blocking buffer as the primary antibody incubation buffer

I hope this is helpful. Please contact me again if you have any further questions.

Thank you for your enquiry.

I am sorry to hear this antibody is giving you trouble.

I just have a few additional questions:

1. Please describe the secondary antibody/detection system a bit further, including dilutions if appropriate.

2. Have you tried any other dilutions of ab5690?

3. Have you used any other antibodies to succesfully stain these same mouse brainsections?

4. With what are the mouse brain sections blocked?

Thanks in advance for the additional information!

According to our records, ab5690 was proving difficult to use in IHC-P and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help resolve this case.

Could you please let me know your PO or order number so that I can find the order in our system and decide whether a free of charge replacement would be an option (if ordered within the last 6 months)?Thank you so much!

If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

I wish you the best of luck with your research and look forward to a reply.

Thank you for your email.

We have 4 rabbit antibodies that work in rat in IHC-P. I have listed them below:

ab16044 Anti-CD3 antibody
https://www.abcam.com/ab16044

ab16669 Anti-CD3 antibody [SP7]
https://www.abcam.com/ab16669

ab13628 Anti-CD3 antibody
https://www.abcam.com/ab13628

In order to help investigate the problems with ab5690 and see if there are any tips I can offer to help, would it be possible to find out a bit more information about your protocol? I have included some questions below. Looking forward to your reply.

Description of the problem (no staining, non-specific or incorrect staining, high background, etc)
Product lot number
Sample preparation (tissue/cell linetested , species, how these have been fixed, any antigen retrieval steps performed, permeabilization steps)
Blocking conditions (buffer used, blocking time/temperature)
Primary antibody conditions (manufacturer/species/target, diluent, dilution used, incubation time/temperature)
Secondary antibody conditions (manufacturer/species/target, diluent, dilution used, incubation time/temperature)
Wash steps (buffer used, time and number of washes)
Detection (manufacturer, detection method used: DAB/fluorescent /etc, any amplification step used)
Controls (positive/negative, no primary)
Any troubleshooting steps you have attempted, how many times you have attempted to run the experiment
Any order information that will help us locate your original order in our system (Abcam order number, PO number, date)