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Cultured T cells from an IL-2-dependent T-cell line (CA1) prepared from human peripheral blood activated with influenza virus.
Our Abpromise guarantee covers the use of ab23 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|IP||Use at an assay dependent concentration.|
|WB||Use a concentration of 5 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 131 kDa).|
|IHC-P||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
ab23 (1 µg/ml) staining CD45RO in human tonsil.
Sections were stained using an automated system (DAKO Autostainer Plus ), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH 6.1. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with hematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab23 (red line). The cells were fixed with 80% methanol (5 minutes) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23, 1 µg/1x106 cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) ab96879 at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"