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Our Abpromise guarantee covers the use of ab27526 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. PubMed: 18156632|
|IHC-FoFr||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||1/200. for 10 minutes at room temperature. Staining of Formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 minutes followed by cooling at room temperature for 20 minutes.|
ChIP analysis using ab27526 binding Notch1 in mouse pancreatic cancer primary cells. Cells were cross-linked for 12 minutes with 1% formaldehyde. Samples were incubated with the primary antibody at a dilution of 1/50 for 8 hours at 4°C. Protein binding was detected using real-time PCR.
Pol II as was used as a positive control and IgG as background control. GAPDH promoter was used as the non-binding control.
ChIP Dilution Buffer:
1.1% Triton X-100,
16.7mM Tris-HCl, pH 8.1,
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 (ab150073) (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma labelling Notch1 with 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.
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 4°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (ab6720) (1/1000) was used as the secondary antibody.
ab27526 staining Notch1 in Mouse brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde and blocked with 3% serum for 30 minutes at 25°C. The sample was incubated with primary antibody (1/50 in 3% Horse serum - ab7484) at 4°C for 15 hours. An undiluted HRP-conjugated Horse anti-rabbit IgG polyclonal was used as the secondary antibody.
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