重组Anti-Cytokeratin 5抗体[SP27] (ab64081)

Flow cytometry overlay histogram showing left A-431 positive cells and right negative MCF7 stained with ab64081 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab64081) (1x 10⁶ in 100μl at 1.0μg/ml (1/2080)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Fluorescence multiplex immunohistochemical analysis of human prostate gland tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-Prostate Specific Antigen (ab76113, red; Opal™690), anti-Cytokeratin 5 (ab236216, green; Opal™520) and anti-Nkx3.1 (ab196020, gray; Opal™570) on human prostate gland tissue. Panel B: anti-Prostate Specific Antigen stained on cytoplasm of luminal cells. Panel C: anti-Cytokeratin 5 stained on basal cells. Panel D: anti-p63 stained on nucleus of luminal cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab76113 (1/2000), ab236216 (1/400) and ab196020 (1/2000) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

Fluorescence multiplex immunohistochemical analysis of human prostate gland tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-p63 (ab124762, magenta; Opal™690), anti-Cytokeratin 5 (ab236216, green; Opal™520) and anti-Prostate Specific Antigen (ab76113, red; Opal™570) on human prostate gland tissue. Panel B: anti-Prostate Specific Antigen stained on luminal cells. Panel C: anti-Cytokeratin 5 stained on cytoplasm of basal cells. Panel D: anti-p63 stained on nucleus of basal cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab124762 (1/5000), ab236216 (1/400), and ab76113 (1/2000) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

Fluorescence multiplex immunohistochemical analysis of human prostate gland tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-Collagen VI (ab271938, magenta; Opal™690), anti-Cytokeratin 5 (ab236216, green; Opal™520) and anti-Prostate Specific Antigen (ab76113, red; Opal™570) on human prostate gland tissue. Panel B: anti-Prostate Specific Antigen stained on luminal cells. Panel C: anti-Cytokeratin 5 stained on basal cells. Panel D: anti-Collagen VI stained on stroma. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab271938 (1/500), ab236216 (1/400), and ab76113 (1/2000) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat skin tissue sections labeling Cytokeratin 5 with ab64081 at 1/100 dilution (2.46 µg/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Cytoplasmic staining on rat skin, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64081 for 30 mins at room temperature.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse skin tissue sections labeling Cytokeratin 5 with ab64081 at 1:100 dilution (2.46 ?g/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Cytoplasmic staining on mouse skin, performed on a Leica Biosystems BOND^TM RX instrument
The section was incubated with ab64081 for 30 mins at room temperature.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung squamous cell cancer tissue sections labeling Cytokeratin 5 with ab64081 at 1/100 dilution (2.46 µg/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Cytoplasmic staining on tumor cells of human lung squamous cell cancer, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64081 for 30 mins at room temperature.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human prostate cancer
tissue sections labeling Cytokeratin 5 with ab64081 at 1/100 dilution (2.46 µg/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on human prostate cancer, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64081 for 30 mins at room temperature.

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Immunohistochemistry (Frozen) analysis of mouse skin tissue section labeling Cytokeratin 5 with purified ab64081 at 1/30 (8.2 µg/ml). Sections were fixed in 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Antigen retrieval was Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunocytochemistry/ Immunofluorescence analysis of A431 (human epidermoid carcinoma epithelial cell) cells labeling Cytokeratin 5 with purified ab64081 at 1/100 (2.5 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunohistochemistry (Frozen) analysis of rat skin tissue section labeling Cytokeratin 5 with purified ab64081 at 1/30 (8.2 µg/ml). Sections were fixed in 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Antigen retrieval was Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Flow cytometric analysis of A431 (human epidermoid carcinoma cell line) cell line labeling Cytokeratin 5 with ab64081 at 1/100 dilution (green) compared with a negative control of rabbit IgG (blue).

Formalin-fixed, paraffin-embedded human prostate tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human thymus tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human skin tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human pancreatic adenocarcinoma tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human lung tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human lung squamous cell carcinoma tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human breast tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human cervical squamous cell carcinoma tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human bladder tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human bladder transitional cell carcinoma tissue stained for Cytokeratin 5 using ab64081 at 1/100 dilution in immunohistochemical analysis.