Immunohistochemistry (Formalin-fixed paraffin-embedded sections) - Notch1 antibody (ab27526)

Staining of Ovarian carcinoma using ab27526

Immunohistochemistry (Frozen sections) - Notch1 antibody (ab27526)

ab27526 staining Notch1 in mouse neural tube by immunohistochemistry (Frozen). Sections were paraformaldhyde fixed prior to blocking in 20% serum for 16 hours at 25°C and then incubated with ab27526 for 16 hours at 25°C. A Cy3 conjugated donkey polyclonal to rabbit Ig was used as the secondary antibody. Blue staining is nuclear staining with DAPI and the specific Notch signal is red.

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence - Notch1 antibody (ab27526)

ab27526 staining Notch 1 in Mouse pancreatic cancer primary cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone/methanol (1:1) and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in PBS) for 4 hours. An Alexa Fluor^®488-conjugated Donkey anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI.

This image is courtesy of an anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Notch1 antibody (ab27526)

ab27526 staining Notch 1 in Human pancreatic neoplastic lesion tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in antigen unmasking solution. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4ºC. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

Western blot - Notch1 antibody (ab27526)

Anti-Notch1 antibody - ChIP Grade (ab27526) at 1/500 dilution (in TBST for 8 hours at 4°C) + Whole cell lysate of human MiaPaCa cells at 50 µg

Secondary
An HRP-conjugated Donkey anti-rabbit IgG polyclonal at 1/5000 dilution
developed using the ECL technique

Performed under non-reducing conditions.

Observed band size : 120,250 kDa (why is the actual band size different from the predicted?)
Additional bands at : 75 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 10 minutes

Blocking Step: 5% Milk for 1 hour at room temperature.

This image is courtesy of an anonymous Abreview