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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Oct4 aa 250-350.
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
A trial size is available to purchase for this antibody.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Alternative versions available:
Our Abpromise guarantee covers the use of ab109183 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Predicted molecular weight: 39 kDa.|
|IP||1/10 - 1/100.|
|IHC-P||1/500 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|Flow Cyt||1/10 - 1/100. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.|
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: POU5F1 (KO) knockout HAP1 whole cell lysate (20 µg)
Lanes 1 - 2: Merged signal (red and green). Green - ab109183 observed at 38 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab109183 was shown to specifically react with POU5F1 (KO) when POU5F1 (KO) knockout samples were used. Wild-type and POU5F1 (KO) knockout samples were subjected to SDS-PAGE. Ab109183 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 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.
ab109183, at 1:500 dilution, staining Oct4 in paraffin-embedded Human ovarian dysgerminoma by Immunohistochemistry.
Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Oct4 with ab109183 at 1/60 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
ab109183 at 1/50 immunoprecipitating Oct4 in NCCIT (human pluripotent embryonal carcinoma) whole cell lysate observed at 39 KDa (lanes 1 and 2).
Lane 1 (input): NCCIT whole cell lysate, 10μg
Lane 2 (+): ab109183 + NCCIT whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109183 in NCCIT whole cell lysate
For western blotting, ab109183 at 1/1000 dilution and ab131366 VeriBlot for IP (HRP) was used as the secondary antibody at 1/10000.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Flow cytometry analysis of NCCIT (human pluripotent embryonal carcinoma cells labelling Oct4 (pink) with ab109183 at dilution of 1/100. The secondary antibody used was goat anti rabbit IgG (FITC) at dilution of 1/150. Cells were fixed with 2% paraformaldehyde. Isotype control antibody used was rabbit monoclonal IgG (green).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"