FITC Anti-TLR2 抗体 [T2.5] (ab59711)

Thank you for your phone call.

The free of charge replacement antibody you requested, ab24192, will ship on order ***. This antibody has not been validated for use on frozen sections, but may be suitable for that purpose. The antibody may require antigen retrieval for longer PFA fixations. If you are unable to use a shorter fixation method, the attached reference provides details about how to perform heat-mediated antigen retrieval on frozen tissue sections. I hope this helps, please let me know if you need any additional information or assistance.

I have found another reference that is more encouraging:

TLR2 is constitutively expressed within the kidney and participates in ischemic renal injury through both MyD88-dependent and -independent pathways.
J Immunol. 2007 May 15;178(10):6252-8.

http://www.jimmunol.org/content/178/10/6252.full

The mouse tissue was PFA-fixed and embedded frozen in OCT, and it does not appear any antigen retrieval was necessary. It is not clear if the antibody was directly conjugated, or if a secondary antibody was applied.

Confocal imaging and analysis

To prepare tissue for confocal imaging, experimental and control groups of WT and TLR2-deficient mice were sacrificed and perfused in situ with 2% paraformaldehyde. Immediately following the perfusion, the kidneys were dissected, removed, and fixed in a solution of 4% paraformaldehyde overnight. The next morning, the kidneys were placed through a sucrose gradient and then frozen in OCT medium (Sakura). Sections (6 μm thick) were stained with anti-TLR2... (see figure 1).

I found this reference and others by searching via Google Scholar, using the keywords "T2.5" (the clone ID) and TLR2, and formalin or paraformaldehyde.

I hope this helps. Please let me know if you have any questions.

Thank you for contacting us. So far, the one reference I have found that used this antibody with frozen tissue sections used acetone as the fixative, not PFA.

Kidneys were snap frozen in liquid nitrogen and stored at –70°C prior to use. For histological examination of TLR2, kidneys were cut into 6-μm sections, fixed for 10 minutes with ice-cold acetone, and air dried. The sections were incubated in 5% normal goat serum (DakoCytomation) for 10 minutes and then exposed overnight to anti-TLR2 mAb T2.5 in PBS.

Leemans, J et al; Renal-associated TLR2 mediates ischemia/reperfusion injury in the kidney. J Clin Invest 2005 115: 2894

I have asked for more data from our source for this clone and will forward this when I receive it. Can you please confirm the species of your samples? I am assuming mouse embryo.