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corresponding to Human CD168 aa 1-100.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Our Abpromise guarantee covers the use of ab124729 in the following tested applications.
|WB||1/1000. Predicted molecular weight: 84 kDa.
For unpurified use at 1/1000 - 1/10000.
The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.
|IP||1/10 - 1/100.|
For unpurified use at 1/50 - 1/100.
Antigen retrieval is recommended
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human gastric carcinoma tissue sections labeling CD168 with purified ab124729 at 1:250 dilution (7.7 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
ab124729 (purified) at 1:100 dilution (2ug) immunoprecipitating CD168 in MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate.
Lane 1 (input): MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab124729 & MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124729 in MCF-7 (Human breast adenocarcinoma epithelial cell) 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.
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Empty knockout HAP1 whole cell lysate (20 µg)
Lane 3: CD168 whole cell lysate (20 µg)
Lanes 1 - 3: Merged signal (red and green). Green - ab124729 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab124729 was shown to specifically react with CD168 when CD168 knockout samples were used. Wild-type and Empty knockout samples were subjected to SDS-PAGE. Ab124729 and ab9484 (Mouse anti GAPDH 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 1/20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemical analysis of paraffin-embedded human testis tissue using unpurified ab124729 at 1/50 - 1/100 dilution