Mouse, Human Predicted to work with:
Rat, Rabbit, Horse, Cow, Chimpanzee, Macaque monkey, Gorilla, Chinese hamster, Orangutan
Synthetic peptide corresponding to Mouse Rubicon aa 750-850 conjugated to Keyhole Limpet Haemocyanin (KLH). Database link: Q80U62
This antibody gave a positive signal in the following tissue lysates: Mouse Liver; Mouse Kidney; Human Liver; Human Kidney.
This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
ICC/IF image of ab156052 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab156052 at 10µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h.
ab50533 (Mouse monoclonal [Rab7-117] to RAB7) was also incubated along with ab156052 to demonstrate antigen co-localization. Signal detected using Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed (ab150120) (pseudo-colored red) DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Western blot - Anti-Rubicon antibody (ab156052)
All lanes : Anti-Rubicon antibody (ab156052) at 1 µg/ml
Lane 1 : Liver (Mouse) Tissue Lysate Lane 2 : Kidney (Mouse) Tissue Lysate Lane 3 : Human liver tissue lysate - total protein (ab29889) Lane 4 : Human kidney tissue lysate - total protein (ab30203)
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 106 kDa Observed band size : 106 kDa Additional bands at : 42 kDa (possible non-specific binding),48 kDa (possible non-specific binding).
Exposure time : 30 seconds
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab156052 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.