Anti-Iba1 antibody (ab5076)

Thank you for those last details, these confirm that your protocol is excellent and should work with ab5076. Looking at the shipping conditions of your order I found that Fedex had left the package at the front door. I therefore wonder how long the package was left there, maybe this caused the antibody to be damaged. We sell a lot of this antibody and have not had any other complaints so I think the problem is due to a damaged vial. Would you like a refund, or a replacement vial? I look forward to hearing from you to resolve this matter,

I'm very sorry to hear you are experiencing problems with ab5076. The fact that another antibody worked well in your samples shows that levels of Iba1 are sufficiently high to be detected and that the problem lies with the use of the Abcam antibody or the antibody. I think your protocol is very good and should work with ab5076. I would appreciate if you could clarify what incubation time you have tried with the primary, what dilution buffer was used and if you can provide your order details so I can look into a shipping delay suggesting the antibody was damaged. I am sorry for the delay and thank you in advance for providing this information,

Thank you for your contacting Abcam and sharing your results with us. We would like to emphasize that this antibody is a Fast-track product and it is not fully characterised yet. Originally, it has been tested in peptide ELISA but not tested in other applications yet. Western Blot: Approx. 25-30 kDa band observed in U937 and Human Brain lysates at 0.5-1ug of primary. We encourage customers to feed their results back to us, whether positive or negative, and make all information about each of our products available to researchers. We have received one very good feedback message from one of our customers saying that ab5076 worked successfully for IHC. We would perhaps suggest that you should take a look at this review published on-line on the product datasheet. Try to fix the cells in ice-cold acetone (for 5 or 10 min) or ethanol and make sure that you block the non-specific binding sites with 1-3% BSA before the staining. You may also need to optimise the concentration of the antibody, try to test the antibody at 1/100 dilution and then adjust the concentration according to the results. We hope this will be useful. Should you still have problem with this antibody, then please do not hesitate to contact us again.

Thank you for sending me the details of your protocol. All of the originator's results with this antibody are already stated on the online datasheet. They tested ab5076 with U937 and Human Brain lysates and saw a band at approximately 25-30 kDa. It is noted on the datasheet that "currently we cannot find an explanation in the literature for the band we observe given the predicted size of approx 17KDa according to NP_001614. We would appreciate any feedback from people in the field - have any results been reported with other antibodies/lysates? Have any splice variants/modified forms been reported?" Thus, we do not have any more information and cannot recommend a known positive or negative control. Because this antibody is not fully characterized, it was released as a fast-track antibody, and we are very eager for feedback. Other than what you have reported, we have only heard from a customer who tried it in immunofluorescence studies on frozen PFA fixed mouse brain sections but failed to detect any staining of microglial cells. I'm afraid that at this time I can only suggest perhaps trying different samples in order to see if you consistently see bands at the same size. If you have any more questions, please do contact us again.