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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human beta Actin.
(Peptide available as ab13772.)
This antibody has been designed for use as a loading control and is ideal for this purpose.
Our Abpromise guarantee covers the use of ab8229 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.|
|WB||1/500. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human beta Actin peptide (ab13772).|
IHC image of beta Actin staining in Human normal colon formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8229, 10µg/ml overnight at +4°C. An HRP-conjugated secondary (Ab6885, Donkey polyclonal to Goat IgG - H&L (HRP) 1/250 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
This blot was produced using a 10% 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 membrane was then blocked for an hour using 5% Bovine Serum Albumin (lanes 1-4) or 3% milk (lanes 5-8) before being incubated with ab8229 overnight at 4°C. Antibody binding was detected using an anti-Goat antibody conjugated to HRP, and visualised using ECL development solution. A non-specific band at 50-kDa is observed on western blots of some of our batches of ab8229. We have found that milk blocking is more effective than BSA blocking in eliminating this non-specific binding.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"