Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)

Paraffin-embedded sections of hippocampus dentate gyrus stained for  BrdU-positive cells. The method is an indirect immunohistochemical technique: the primary antibody is ab6326 diluted 1/100 in PBS and the secondary antibody is an Alexa 488-conjugated anti-rat made in goat diluted 1/200.

Immunohistochemistry (Frozen sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)

ab6326 used in double labeling on mouse sciatic nerve tissue (5µm) sections. This picture was kindly submitted by Dr Ulrich Hengst as part of his review on this product.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)

ab6326 at 1/200 staining mouse E14.5 embryonic telencephalon tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retireval step was performed. The tissue was then stained with the antibody for 12 hours. A Texas Red conjugated donkey anti-rat antibody was used as the secondary.

This image is courtesy of an Abreview submitted by Dr Bing Lang

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)

ab6326 staining mouse adult epidermis tissue sections by IHC-P.  Sections were PFA fixed and subjected to heat mediated antigen retrieval in citrate buffer prior to blocking in 10% serum for 1 hr at RT.  The primary antibody was diluted 1/200 and incubated with the sample for 24 hrs at 4°C.  A biotinylated goat anti-rat antibody was used as the secondary followed by a streptavidin-Cy3®.

This image is courtesy of an anonymous Abreview

Immunocytochemistry - BrdU antibody [BU1/75 (ICR1)] (ab6326)

ab6326 staining cultured cells of rat brain tissue by ICC.  The sample was PFA fixed and permeabilized in 1M HCl prior to blocking with 5% serum for 1 hour at 25°C.  The primary antibody was diluted 1/500 and incubated with the sample for 16 hours at 25°C.  A biotinylated rabbit anti-rat IgG antibody, diluted 1/200, was used as the secondary.

This image is courtesy of an anonymous Abreview

Immunohistochemistry (Frozen sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)

ab6326 staining BrdU in mouse brain tissue sections by IHC-Fr (paraformaldehyde-fixed frozen sections). Tissue samples were fixed with paraformaldehyde; permeabilized win 0.3% Triton X-100 and blocked with 5% Serum for 2 hours at 4ºC. Before permeabilization samples were pretreated with 2N HCl at RT for 30 min and washed 3 times. The sample was incubated with primary antibody (1/200) at 4ºC for 12 hours. An Alexa Fluor^® 488-conjugated Goat polyclonal to rat IgG (1/500) was used as secondary antibody. BrdU staining shown in green and NewN staining showin in red.

This image is courtesy of an anonymous Abreview

Flow Cytometry - Anti-BrdU antibody [BU1/75 (ICR1)] (ab6326)

ab6326 staining BrdU in HeLa cells by Flow Cytometry. Cells were incubated with 10 µM BrdU for 30 minutes prior to being harvested with 1X trypsin-EDTA, washed twice in PBS containing 1% BSA, and fixed in 70% ethanol (added drop-wise) for at least 30 minutes on ice. Once fixed, pellets were acid denatured with HCl/Triton X-100 for 30 minutes at room temperature and then neutralised with sodium tetraborate.
Pelleted cells were re-suspended in Tween/BSA/PBS to which primary antibody was then added (0.1 µg in 0.5% Tween 20 (v/v) plus 1% BSA in PBSA) and incubated for 30 minutes at room temperature. Secondary Alexa Fluor®488-conjugated Goat anti-Rat IgG (H+L) was used at 1/500 and incubated for 30 minutes at room temperature in the dark. Cells were pelleted once more and resuspended in PBS containing 5 µg/mL propidium iodide.
Gating Strategy: Based on forward and side scatter, cells were gated into the region used for analysis. This was done by applying a large circle to accommodate the normal cells in an untreated control.

This image is courtesy of an anonymous Abreview