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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Cytokeratin 8+18 aa 300-400 (C terminal). The exact sequence is proprietary.
Database link: P05787
This product is a recombinant rabbit monoclonal antibody.
A trial size is available to purchase for this antibody.
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
Our Abpromise guarantee covers the use of ab53280 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/25000 - 1/50000. Detects a band of approximately 52 kDa (predicted molecular weight: 54 kDa).|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||1/100 - 1/500.|
Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School
Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)
Fix in 3%PFA in PBS for 30 min at RTIncubate in 7.5% sucrose-PBS for 3h at RTIncubate in 15% sucrose-PBS at 4 degree Celsius overnightEmbed the EBs in tissue-Tek OCT compoundCut frozen sections to 4-20 µm thickness
Primary antibody 1: Rabbit anti cytokeratin 8 (ab53280), 1:100
Primary antibody 2: Rat anti-perlecan, 1:100
Secondary antibody 1: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081), 1:200
Secondary antibody 2: Goat polyclonal Secondary Antibody to Rat IgG - H&L (Cy5®) pre-adsorbed (ab150081), 1:200
Nuclei were counterstained with DAPI
ab53280 staining Cytokeratin 8+18 in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling Cytokeratin 8+18 with purified ab53280 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised by 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Capture ab: 1/20 dilution (ab133272, 2μg in 0.35mg lysates)
Primary ab for WB:
- Panel A: CK-18 (ab133272,1/1000)
- Panel B: CK-8 (ab53280,1/1000)
Secondary ab: VeriBlot for IP secondary antibody (HRP) (ab131366)
Blocking and dilution buffer: 5% NFDM/TBST
Lane 1 (Input): HeLa (human cervix adenocarcinoma) whole cell lysate 10ug
Lane 2 (+): HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133272 in HeLa (human cervix adenocarcinoma) whole cell lysate
- Panel A: 30 seconds
- Panel B: 1 seconds
Overlay histogram showing HeLa cells stained with ab53280 (red line). The cells were fixed with 2% PFA (room temperature, 30 min) and then permeabilized with 1% FACS permeabilizing solution for 30 min. The cells were then incubated in 3% FBS in 1X PBS followed by the antibody (ab53280, 1/20 dilution) for 1 hour at room temperature. The cells were then incubated for 30 min at room temperature with the secondary antibody. An isotype control antibody (black line) was used and an unlabelled sample (blue line) was also used as a control.