Anti-Campylobacter jejuni抗体[BGN/2E10] (ab54125)

This antibody has been used in the following publication:

Haddock G et al. Campylobacter jejuni 81-176 forms distinct microcolonies on in vitro-infected human small intestinal tissue prior to biofilm formation. Microbiology 156:3079-84 (2010).

The protocol used was as follows:

Human ileal sections of 4 μm thickness were mounted onto slides and allowed to dry before placing in a 37 °C oven overnight. Slides were marked with the antibody and date, dewaxed in xylene and rinsed in 100 % alcohol then 95 % alcohol. Slides were then placed in 3 % hydrogen peroxide in methanol for 20 min at room temperature. After washing in water, antigen retrieval was performed by placing the slides in an autoclave for 10 min in citrate buffer, pH 6, at 121 °C. Slides were left to cool to 50 °C before the transfer of slides to a staining chamber. Slides were washed in PBS/Tween-20 twice before applying 25 % normal goat serum for 30 min at room temperature. Antibody against C. jejuni was added to the slides overnight at 4 °C (100 μl, 1 : 600 mouse monoclonal, BGN/2E10, 1 mg ml−1, Abcam ab54125).

You could therefore do a trial run with this antibody at a dilution of 1:600 overnight at 4ºC and then optimise accordingly.

ab54125 wurde von uns nur mit Aceton-fixiertem Gewebe getestet und wir haben leider auch sonst keinerlei Anhaltspunkte (Rückmeldung von Kunden, etc.), dass er mit PFA-fixierten Geweben funktioniert.

Da auch der andere Antikörper nicht mit PFA funktioniert hat, schien es nicht per se am Antikörper zu liegen sondern eher daraufhin zu deuten, dass dieses Zielprotein nicht geeignet ist für einen PFA Fixierung.