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Our Abpromise guarantee covers the use of ab48187 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/2000. Predicted molecular weight: 110 kDa.
Block with 3-5% BSA.
|IHC-P||1/10 - 1/500.|
|ELISA||1/100 - 1/2000.|
|IP||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 20477942|
The blots were produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membranes were blocked for an hour. Membrane 2 was incubated with alkaline phosphatase (AP; 100 U per mL) for one hour, whilst membrane 1 was treated with AP-buffer only, before being incubated with ab48187 (rabbit anti-IRE1 antibody diluted 1:2000) and loading control ab125247 (mouse anti-GAPDH antibody; diluted 1:10,000) for 24 hours at 4°C. Antibody binding was detected using infrared (IR)-labelled goat anti-rabbit (green) antibody and IR-labelled goat anti-mouse (red) at 1:10,000 dilutions for 1 hour at room temperature before imaging.
Serially diluted ab48187 was bound to immobilised Phospho peptide (133861) - or Control peptide (1 microgram x mL-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 50000 times) and signal was developed with TMB substrate.