Anti-F4/80 antibody [CI:A3-1] (ab6640)

I would like to answer those questions.
1. For antigen retrieval: cooking in citrate buffer for 20 min, leave in citrate buffer for 10 min and cool by keeping it outside for 5 min.
2. The blocking buffer has also been reduced to 3% BSA. But no use.
3. Regarding Rat AP-polymer kit, I don't know which information do you need. Because we follow the kit protocol.
4. We have stained our slides with CD3, CD8 and other antibodies also by following the same protocol and just this one is not working.

ANSWER:

I will issue you replacement or credit note if you wish, however, i would just like to make some comments:

I was asking about the Rat AP-polymer kit because we usually like to check the compatibility with our products as a few cause trouble sometimes and we than can simply recommend a different one.

Please keep in mind that you cannot compare two different antibodies against two different targets with each other. Every antibody needs its individual optimization.


Thank you again for your time and cooperation. Please let me know on how you would to proceed.

Thank you for responding so quickly. We will either be using formalin fixed, paraffin embedded adipose tissues if possible, if not, frozen (OCT) adipose tissue will be used. I have always been under the impression that antigen retrieval is the major difference between these, in which case we would prefer to use the formalin fixed tissue. The tissues will receive no other treatment. Of the antibodies available do you have a suggestion of which would be most effective in this tissue? And then we can decide the proper conjugated secondary. Thanks

ANSWER:

Thank you for your enquiry and for this further information.

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 CD11c, F4/80 or Mannose Receptor antibodies tested in the species and application you are using should be suitable for your applications.

Also, 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.

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).

Using these criteria I can recommend ab33483 anti-CD112c, ab6640 anti-F4/80 and ab8918 anti- Mannose receptor antibodies. These products are all guaranteed to work in numerous applications and have been used in 14, 53 and 9 publications respectively.

As these products are raised in Hamster, Rat and Mouse you should experience few issues in using all three together however I would suggest that you try to find secondary antibodies which are isotype specific e.g. Hamster IgG, Rat IgG2b and Mouse IgG1. Should you wish for assistance in finding the correct secondary antibodies, I would be happy to help. To do this I would need to know your conjugation requirements such as which enzymes, dyes or fluorescent molecules you prefer. I would also like to let you know that we are currently running a promotion for 25% off all secondary antibodies.

I have posted links to these antibodies below:

CD11c http://www.abcam.com/CD11c-antibody-N418-ab33483.html

F4/80http://www.abcam.com/F4-80-antibody-CI-A3-1-ab6640.html

Mannose Receptorhttp://www.abcam.com/Mannose-Receptor-antibody-15-2-ab8918.html


I look forward to hearing from you. Should you have any further questions please do not hesitate to ask.

I am somewhat new to immunofluorescence but am attempting to identify M1/M2 polarized macrophage markers in mouse adipose tissue utilizing this method. I have seen Abcam referenced many times for my desired markers in publication so I wanted to come here first. I am looking to stain for markers F4/80, CD11c, and CD206. From what I can tell from your website, you have a few antibodies that react with mouse tissue for all of these (ab33483 and ab11029 for CD11c)(ab6640, ab16911 and ab60343 for F4/80) and for the CD206, anti-mannose receptor antibodies come up (ab8918 and ab64693 are for mouse). I am assuming then that anti-mannose receptor antibodies refer to the C type I mannose receptor. Furthermore, can you advise me on what I will need to order, including mounting medium and such, to stain for these markers in mouse adipose tissues and visualize them using fluorescence? Thank you very much for your assistance.

ANSWER:

Thank you for contacting us.

I would be more than happy to assist you in picking the corrct antibodies and reagents to suit your needs. To begin I only need to know a little more about the sample preparation which you will be performing. It is most important to understand any treatment that the tissue will recieve prior to staining.If you are using tissue will this be formalin fixed paraffin embedded, or will you be using another technique and fixative? If you are using monolayer cells what type of fixation will you use?

I have included a few pdfs one of which details our general IHC/ICC technique (immunostaining at a glance) as well as two more in depth ones.

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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ab6640 Anti-F4/80 antibody [CI:A3-1]doesn't work: new lot failed, but old one works fine with same protocol.

ANSWER:

Thank you for confirming these details and for your cooperation.


I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number XXXXX. It should arrive Monday, 14. of May at the Apotheke des Universitatsklinikums.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

IHC-Fr with mouse ear and lymphnodes
acetone fixation
staining: specific + bright specks (non-specific)
no primary crtl: clean

Ab: 1/1000 RT 1h
block: goat serum
2nd: Alexa555 1/400

suggested: less primary and secondary Ab, block with BSA, block longer

ANSWER:

Thank you for contacting us.

As we discussed over the phone, I'd suggest to try the following to get rid of the specks you see in your images:

1) use less primary antibody
2) use lesssecondary antibody
3)block with 5% or 10% BSA
4) tryblocking longer (e.g. 2-3 h at RT or overnight at 4 degree)

Please let me know if these tips are of help or not.

I wish you good luck and look forward to hear back from you.

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