Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody (ab59400)

ab59400 staining Cytokeratin 8 in c57b6 mouse prostate tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using citric acid. Samples were then blocked using 3% BSA for 1 hour at 25°C and then incubated with ab59400 at a 1/100 dilution for 18 hours at 4°C. The secondary used was a biotin conjugated goat anti-rabbit polyclonal used at a 1/200 dilution.

Image courtesy of an anonymous Abreview.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Cytokeratin 8 antibody (ab59400)

Immunohistochemical analysis of paraffin embedded human colon carcinoma tissue using ab59400 at 1/50 dilution, in the presence (right) and absence (left) of immunising peptide. Then, a polymer secondary antibody was used for detection.

Western blot - Cytokeratin 8 antibody (ab59400)

All lanes : Anti-Cytokeratin 8 antibody (ab59400) at 1/500 dilution

Lane 1 : EGF treated (200ng/ml, 30mins) 293 cell extracts
Lane 2 : EGF treated (200ng/ml, 30mins) 293 cell extracts with immunising peptide


Predicted band size : 54 kDa
Observed band size : 54 kDa

Immunocytochemistry/ Immunofluorescence - Cytokeratin 8 antibody (ab59400)

ICC/IF image of ab59400 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab59400, 1µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Cytokeratin 8 antibody (ab59400)

ab59400 staining Cytokeratin 8 in Human breast cancer cells xenografted into nude mice by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). The sections were fixed in formalin and subjected to heat-mediated antigen retrieval in citrate buffer (0.1M Sodium Citrate) prior to blocking with 5% serum for 1 hour at 4ºC. The primary antibody was diluted 1/200 in 5% goat serum in PBS and incubated with the sample for 12 hours at 4ºC. A Biotin-conjugated Goat anti-Rabbit polyclonal was used as the secondary antibody, diluted 1/1000.

This image is courtesy of an Abreview provided by Megha Rajaram.

Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 8 antibody (ab59400)

ab59400 staining Cytokeratin 8 in murine mammary gland duct tissue by Immunohistochemistry (Frozen sections).

Tissue was fixed in paraformaldehyde, permeabilized using 0.1% Triton X-100, blocked with 1% BSA for 1 hour at 23°C and then incubated with ab59400 at a 1/250 dilution for 12 hours at 4°C. The secondary used was an Alexa-Fluor 568 conjugated goat anti-rabbit polyclonal used at a 1/1000 dilution.

Image courtesy of an anonymous Abreview.