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Synthetic peptide within Human PDCD4 (N terminal). The exact sequence is proprietary.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
This product is a recombinant rabbit monoclonal antibody.
Our Abpromise guarantee covers the use of ab80590 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/20000 - 1/100000. Predicted molecular weight: 52 kDa.|
|ICC||1/100 - 1/250.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||Use at an assay dependent concentration.|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: PDCD4 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab80590 observed at 52 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab80590 was shown to specifically react with PDCD4 in wild-type HAP1 cells as signal was lost in PDCD4 knockout cells. Wild-type and PDCD4 knockout samples were subjected to SDS-PAGE. ab80590 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 20000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) labelling with ab80590 at a dilution of 1:100 dilution (0.34 µg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) (1:1000 dilution (2 µg/ml)) was used as the secondary antibody. The cells were co-stained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Nuclei counterstained with DAPI (blue). Control: PBS instead of the primary antibody.
Lane 1 (input): HeLa(Human cervix adenocarcinoma epithelial cell) whole cell lysate, 10µg
Lane 2 (+): ab80590 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab80590 in HeLa whole cell lysate
Ab80590 (Purified) at 1:500 dilution (1.184 µg/ml) immunoprecipitating PDCD4 in HeLa whole cell lysate. For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST .