Anti-Iba1 抗体 (ab108539)

Thank you for contacting Abcam.

So that I can better help you with this, would you be able to provide me with a copy of the image that you are getting and also the protocol that you are using, in particular could you include the following information:

1 - What sample type are you using?

2 - How much sample are you loading?

3- What type of blocking agent are you using?

4- What primary antibody concentrations have you tried and how long do you incubate for?

5 - How long do you boil your samples for?

I look forward to your reply and helping you resolve this issue.

Thank you for contacting us.

I can confirm that antibody ab108539 could be used for detection of Iba1 in microglia cells of mouse brain. This assumption is however based on the expression of this protein in mouse microglia cells; we unfortunately haven't tried this in our lab however it would work.

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 taking the time to contact us. I am sorry to hearthe customer hashad difficulty obtaining satisfactory results from these threeantibodies.

I would like to reassure you that 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.

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 also provide images which would help us to assess the results.

Please note Ihave sent the questionnaire for IHC and ICC. If the customer has used another application, please let me know and I can send the correct form so we can obtain the relevant information required.

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



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

Here is a quite detailed protocol for performing paraffin embedded immunohistochemical staining, which has the buffer composition for Tris-EDTA pH 9 which I suggested trying: https://www.abcam.com/ps/pdf/protocols/ihc_p.pdf Tris-EDTA buffer: 10 mM Tris Base, 1 mM EDTA, 0.05% Tween 20, pH9.0 Its quite a useful guide, especially when troubleshooting for a new experiment. I hope it is of help to you. Have a nice weekend.

Thank you for contacting us again. As discussed over the phone, I have arranged for the free of charge replacement with ab5076 to be delivered to you at the following address (taken from your previous order): The new order number is: This should be with you tomorrow. I hope this one works better for you. Although your collegues have said that they have not required any antigen retrieval, several of our customers have used citrate mediated antigen retrieval so you may need to optimise when using your particular samples. I was also wrong to say that this antibody has not yet been tested in mouse samples, several of our customers have used it for IHC with mouse tissue and I will amend this information on the datasheet. As discussed I have also attached a questionnaire which it would be very helpful if you could fill out. This will allow me to investigate this case internally and initiate any additional testing where necessary. I look forward to receiving your reply.

Thank you for your enquiry. I would like to reassure you that our antibodies have all been tested successfully and would be covered by our guarantee in the applications listed on the datasheets. Therefore, any Iba1 antibody tested in the species and application you are using should be suitable for your applications. I can confirm that both ab5076 and ab108539 have been tested and guaranteed for IHC-P. Although we produce many in-house antibodies, we obtain other antibodies from a wide range of sources. Therefore, comparison experiments will not often have been done. If you wish to be more certain, I can suggest choosing the antibody that has been best characterized. By this I mean the antibody that has been tested in the most applications and has the most data available (on the datasheet). Regarding the secondary antibody, ab6884 Donkey polyclonal Secondary Antibody to Goat IgG (Biotin) is tested and guaranteed in IHC-P. As an anti-goat antibody, it would be suitable for use with Anti-Iba1 antibody (ab5076), as this primary antibody is raised in goat. With regards to ab108539 Anti-Iba1 antibody, this is a rabbit polyclonal, and therefore requires an anti-rabbit secondary antibody and ab6884 would not be a suitable secondary to use with ab108539.. I hope this information will be helpful to you. Should you have any further questions regarding your search and selection of products, please do not hesitate to ask.