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Synthetic peptide corresponding to Human NOTCH3 aa 2300 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).
(Peptide available as
Our Abpromise guarantee covers the use of ab23426 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 97,280 kDa (predicted molecular weight: 244 kDa).|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. PubMed: 19965627|
|ICC/IF||Use at an assay dependent concentration. PubMed: 20680961|
|IHC-FoFr||Use at an assay dependent concentration.|
|ICC||Use at an assay dependent concentration.|
This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab23426 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
The band observed at 97 kDa is thought to correspond to the notch-derived peptide containing the intracellular domain (NICD) of NOTCH3 as described in the literature (PMID:10712431).
ab23426 staining NOTCH3 in mouse embryo tissue (E16.5) by Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections). Tissue underwent fixation in formaldehyde, heat-mediated antigen retreival in citrate buffer pH6.0 and blocking for 15 minutes at 10°C (5 minutes/peroxidase blocking and 10 minutes/protein blocking). The primary antibody was diluted 1/250 and incubated with sample for 15 minutes at 20°C. A HRP-conjugated goat polyclonal to rabbit IgG was used undiluted as the secondary.
ab23426 staining NOTCH3 in human breast cancer tissue sections by immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections). Tissue underwent fixation in paraformaldehyde, heat-mediated antigen retrieval in citrate buffer pH6.0 and blocking for 15 minutes at 20°C (5 minutes for peroxidase blocking and 10 minutes for protein blocks). The primary antibody was diluted 1/250 and incubated with sample for 45 minutes at 20°C. A HRP-conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.
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