Anti-CD3 antibody [CD3-12] (ab11089)

This blot was produced using a 4-12% Bis-tris gel under the MES 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 Licor blocking buffer before being incubated with ab11089 overnight at 4°C. Antibody binding was detected using Goat anti-Rat secondary at a 1:10000 dilution for 1hr at room temperature and then imaged.

IHC image of CD3 staining in a formalin fixed, paraffin embedded normal mouse lymph node tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6). The section was incubated with ab11089 at 1/250 dilution for 15 mins at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was 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.

IHC image of CD3 staining in a formalin fixed, paraffin embedded normal human tonsil tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6). The section was incubated with ab11089 at 1/250 dilution for 15 mins at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was 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.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD3 with ab11089. Heat mediated antigen retrieval was performed using EDTA buffer pH 9.0.

Staining of human peripheral blood lymphocytes with Rat anti Human CD3 (ab11089) by Flow cytometry (FACS).

ab11089 staining CD3 in mouse lymph node tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 15% serum for 60 minutes at 20°C. Antigen retrieval was by heat mediation in NaCitrate (pH 6). Samples were incubated with primary antibody (1/250 in TBS) for 18 hours at 20°C. An Alexa Flour^®conjugated goat anti-rat IgG polyclonal (1/400) was used as the secondary antibody.

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