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Tissue/ cell preparation (Human). (T cell line HPB-ALL).
Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab212319 as a replacement.
Our Abpromise guarantee covers the use of ab1422 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 10 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.|
|IP||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 25254972|
Human peripheral blood lymphocytes stained with ab1422 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab1422, 1μg/1x106 cells) for 30 min at 4°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgM (mu chain) (ab97007) at 1/500 dilution for 30 min at 4°C. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.