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Immunocytochemistry/ Immunofluorescence - Anti-DDX19A antibody (ab108462)This image is courtesy of an Abreview submitted by Dr Kirk McManus
ab108462 staining HeLa cells by ICC/IF. The cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X100 in PBS. The cells were then stained with ab108462 at 1/200 in PBS for 1h at 22°C. An Alexa Fluro 488 goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody. Nuclei are stained in red with DAPI. ab108462 produces the expected nuclear membrane-associated staining pattern.
All lanes : Anti-DDX19A antibody (ab108462) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 54 kDa Observed band size : 54 kDa Additional bands at : 60 kDa (possible non-specific binding).
Exposure time : 90 seconds
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 before being incubated with ab108462 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.