IHC image of Integrin alpha 2 staining in a section of formalin-fixed paraffin-embedded normal human colon*, performed on a Leica BOND™. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab209944, 1/50 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

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.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

All lanes : HRP Anti-Integrin alpha 2 antibody [EPR17338] - C-terminal (ab209944) at 1/5000 dilution

Lane 1 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate at 10 µg
Lane 2 : T-47D (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
Lane 3 : C6 (Rat glioma cell line) Whole Cell Lysate at 10 µg
Lane 4 : Brain (Human) Tissue Lysate - fetal normal tissue at 10 µg
Lane 5 : Wild-type HAP1 cell lysate at 20 µg
Lane 6 : Integrin alpha 2 knockout HAP1 cell lysate at 20 µg

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 129 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?


Exposure time: 4 minutes

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 2% Bovine Serum Albumin before being incubated with ab209944 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.