Anti-BrdU antibody [MoBu-1]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [MoBu-1] (AB8039)

Immunohistochemical analysis of parafin-embedded bromodeoxyuridine-labelled chick embryo cells with (MoBu-1) 5-bromodeoxyuridine

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BrdU antibody [MoBu-1] (AB8039)

ICC/IF image of ab8039 stained HeLa cells, both BrdU treated (left image) and normal cells (right image). The cells were 100% methanol fixed (5 min) and then subjected to acid hydrolysis using 2M HCL in 0.1% PBS-Tween for 30 minutes at room temperature to denature the DNA. They were 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 (ab8039, 10μg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM. Positive staining can be seen in the BrdU treated cells, but not in the normal cells, demonstrating specificity for BrdU.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-BrdU antibody [MoBu-1] (AB8039)

Intracellular Flow Cytometry analysis of CEM (human acute lymphoblastic leukemia) cells labelling BrdU with ab8039 at 1μg/mL. Goat anti-mouse IgG was used as the secondary antibody.The individual cell cycle phases (S, G1, G2/M-phase) are indicated on the figure.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [MoBu-1] (AB8039)

IHC image of ab8039 staining, both in normal and BrdU treated rat liver formalin fixed paraffin embedded tissue sections, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8039, 5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.