Anti-BRCA1 antibody [MS110] (ab16780)

Immunocytochemistry/ Immunofluorescence - BRCA1 antibody [MS110] (ab16780)

ab16780 staining BRCA1 in human A431 epidermoid cancer cells by ICC/IF (immunocytochemistry/immunofluorescence). Cells were formaldehyde fixed, permeabilized by Triton X-100 and blocked 5% BSA for 30 minutes at room temperature. The sample was incubated with the primary antibody (1/50 in BSA) for 1 hour. An Alexa Fluor 488^®-conjugated Goat anti-mouse polyclonal (1/50) was used as the secondary.

This image is courtesy of an Abreview submitted by Dr Alejandro Vazquez-Martin

Immunocytochemistry/ Immunofluorescence - BRCA1 antibody [MS110] (ab16780)

ICC/IF image of ab16780 stained MCF7cells. The cells were 4% PFA 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 (ab16780, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) 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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)

IHC image of ab16780 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, 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 ab16780, 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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRCA1 antibody [MS110] (ab16780)

ab16780 staining BRCA1 in Human skin tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded tissue sections). The sections were fixed in formaldehyde and subjected to heat-mediated antigen retrieval in citrate buffer (pH 6.0) prior to blocking with 2% BSA for 1 hour at 22ºC. The primary antibody was diluted 1/50 and incubated with the sample for 20 hours at 4ºC. A Biotin-conjugated goat anti-mouse polyclonal was used as the secondary antibody, diluted 1/800. Antibody was detected by DAB staining.

This image is courtesy of an Anonymous Abreview.