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IHC image of alpha Adducin staining in rat normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab101002, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-alpha Adducin antibody (ab101002)
All lanes : Anti-alpha Adducin antibody (ab101002) at 1 µg/ml (Milk blocking - 5%)
Lane 1 : Kidney (Mouse) Tissue Lysate Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 25 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution Developed using the ECL technique
ab101002 detects a band at 105 kDa, while this differs to its predicted molecular weight of 81 kDa, the banding pattern observed is consistent with what has been described in the literature PMID:9486854.
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% Milk before being incubated with ab101002 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Anti-alpha Adducin antibody (ab101002)参考文献
has not yet been referenced specifically in any publications.