Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)

Thank you for your email. I am sorry to hear that customer have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further suggestions for troubleshooting;

- Please try 5% BSA for blocking with 2 hours incubation
- Heat the samples for 10 minutes 100C
- Try no primary control

It is true that the antibody is out of guarantee period so I am sorry we cannot provide replacement products; we can only provde the troubleshooting suggestions. If however this customer is your loyal customer then you can certainly offer them incentives for keeping them on board.

I hope this information will be helpful. SHould you have any question please do not hesitate to contact me.

Thank you for contacting us. I am sorry to hear these antibodies are not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring 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. 1) About the secondary antibody ab6728: as indicated on the datasheet, many of our customers have reported seeing brown precipitates in the vials. The brown precipitates are very common with HRP conjugated antibodies; we suggest vortexing the vial and using this antibody as normal. However, I would be pleased to replace, if you wish, this antibody with an alternative catalog reference : ab97046, www.abcam.com/ab97046. 2) About the anti-His tag ab5000: to our knowledge this antibody has never been tested for one hour incubation only but overnight at 4ºC. Some antibodies bind better to their target at 4ºC, some other at room temperature. I think it would be worth trying an overnight incubation at 4ºC for this antibody. The blocking solution should not be problem, this antibody works fine in milk as well as in BSA. Please let me know about the results after an overnight incubation at 4ºC. If the antibody is still not working I will be pleased to offer a replacement or a credit note. Thank you for your cooperation. I look forward to hearing from you with your results using ab5000 and with details of how you would like to proceed with ab6728.

Thank you for your message.

I am pleased to help answer your question.

I can confirm that '1 mg at 2mg/ml' means that the vial contains 1 mg of antibody at a concentration of 2 mg.ml. The concentration is 2 mg/ml. (Therefore the vial contains 500 ul).

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Thank you for your reply.

I can confirm that for the HRP conjugated antibodies the approach that Irecommended is the correct one. Spining the vial down will prevent free HRP to bind to un-specifically to samples and therefore avoid increased background.

If the antibody is not purified and therefore also not conjugated then we would recommend gentle vortexing. This could be whole antiserum for example.
In this casethe antibody does contain proteins and other solutes it would not be abnormal to see a little cloudiness or precipitation, especially after being stored at a low temperature and/or for a long period of time. Most of the time bringing the antibody up to room temperature, mixing it well and then diluting it to a working concentration will help incorporate any protein aggregates and precipitation.

I hope this information will explain the different suggestions we gave and I apologise for the confusion.