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Recombinant full length protein, corresponding to amino acids 1-245 of Human TPMT purified from E. coli (NP_000358).
Our Abpromise guarantee covers the use of ab122984 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 at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|WB||1/1000. Predicted molecular weight: 28 kDa.|
|ELISA||Use at an assay dependent concentration.|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: TPMT knockout HAP1 whole cell lysate (20 µg)
Lane 3: HepG2 whole cell lysate (20 µg)
Lanes 1 - 3: Merged signal (red and green). Green - ab122984 observed at 28 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab122984 was shown to recognize TPMT in wild-type HAP1 cells as signal was lost at the expected MW in TPMT knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TPMT knockout samples were subjected to SDS-PAGE. Ab122984 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
The Recombinant Protein (50ng) and Cell lysates (40ug) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with anti-human TPMT antibody (1:1000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.
Lane 1 : Recombinant Protein
Lane 2 : HepG2 cell lysate
Immunocytochemistry/Immunofluorescence analysis of U87MG cells labelling TPMT with ab122984 at 1/100. An Alexa Fluor® 488-conjugated goat anti-mouse IgG was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Flow Cytometry analysis of U87MG cells labelling TPMT with ab122984, staining at 2-5 µg/1x106 cells (red). A 488-conjugated goat anti-mouse IgG was used as the secondary antibody. Black - Isotype control, mouse IgG.
ab122984 has not yet been referenced specifically in any publications.
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