Anti-Oct4 antibody - Embryonic Stem Cell Marker (ab27985)

I am trying to stain cells from lung tissue frozen in OCT compound and
sectioned with a cryostat (6 micron thick). I'm having trouble with the
stain as it's not working after several attempts; the tissue has been fixed
in acetone, but then I found some comments on your website for the product
about fixing in paraformaldehyde (2% solution for about a minute). I tried
a 1:100 dilution of the antibody. Do you have any suggestions that may be
helpful based on the information I've provided?

ANSWER:

Thank you for contacting Abcam.

Based on the information we have for this antibody, it may be that it is not suitable for IHC-Fr on sections. However based on the information you have provided, you could try to increase the primary antibody concentration to a 1/50 dilution and incubate overnight at 4C.

I have also attached the Abcam protocols book that contains more useful information on troubleshooting with IHC-Fr.

We do have a few other Oct4 antibodies that we guarantee to work in IHC-Fr on mouse sections and I have placed links to these below:

http://www.abcam.com/Oct4-antibody-ChIP-Grade-ab19857.html
http://www.abcam.com/Oct4-antibody-ab18976.html



If there is anything else I can help you with, please let me know.

No staining in fixed frozen human tissue sections.

ANSWER:

Thank you for your call today and for letting us know about the trouble with these antibodies.

Please keep me updated about the results with these antibodies after trying the alterations that we discussed. Here is the article that I mentioned about performing heat retrieval on frozen tissue sections-

http://jhc.sagepub.com/content/53/11/1421.full

This article isn't specific for Brdu pre-treatment but it might be a helpful resource for you if you'd prefer to try a heat denaturation before using the HCl method. I've found another article (attached) which will useful for heat denaturation as well.

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.

We ordered the primary antibody Anti-Oct4 antibody - Embryonic Stem Cell Marker (ab27985) a while ago. Now we want to have secondary antibody Rabbit polyclonal to Goat IgG ab28445. But this ab28445 is not available anymore.   Could you please introduce to me another secondary antibody that subtitutes ab28445? I want to use this secondary antibody for both Western Blot and Immunocytochemistry.  

ANSWER:

Thank you for contacting us.

The closest match we have to ab28445 is ab96930. This is an anti-goat IgG, raised in rabbit, conjugated to DyLight® 650, which has an excitation maximum at 654nm and an emission maximum at 673nm. The Alexa Fluor 680 fluorophore of ab28445, on the ohter hand, has an excitation maximum at 680nm and emission at 702nm. Both are in the far red range of the visible light spectrum.

Click here (or use the following: http://www.abcam.com/index.html?datasheet=96930).

Please do not hesitate to contact us if you need any more advice or information.

In the data sheet you specify that the product form is liquid at a concentration of 1mg/ml, but at the beginning of the page it says that the quantity is 100mg, could you tell me the exact volume deliverd for this price?

ANSWER:

As the quantity is 100µg and the concentration 1 mg/ml (1µg/µl) the volume is 100µg/(1µg/µl)= 100ul.