Anti-Ki67 antibody [B56]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse colon. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [B56] (AB279653)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Ki67 with ab279653 at 1/50 (18.72 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nucleolar staining in HeLa cell line ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human colon. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [B56] (AB279653)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized wild-type HeLa (Human cervix adenocarcinoma epithelial cell) cells and KI67 KO HeLa cells labelling Ki67 with ab279653 at 1/50 dilution (18.72 ug/ml), followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nucleolar staining in Parental HeLa cell line and no staining in MKI67 HeLa KO cell line. ab179513 Anti-beta Tubulin rabbit monoclonal antibody at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 dilution was used to counterstain tubulin (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ki67 antibody [B56] (AB279653)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Ki67 with ab279653 at 1/1000 dilution (0.1ug)/ Right compared with a Mouse monoclonal IgG / Left isotype control. A Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human ovarian carcinoma. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling Ki67 with ab279653 at a concentration of 0.1μg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab279653 Anti-Ki67 antibody [B56] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Ki67 with ab279653 at 1/1000 dilution. The section was incubated with ab279653 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Counterstained with DAPI. Secondary antibody only control : Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution1) for 40 mins.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ki67 antibody [B56] (AB279653)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Parental HeLa (Human cervix adenocarcinoma epithelial cells) (Right)/ MKI67 KO HeLa cells (Left) labelling Ki67 with ab279653 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of paraffin-embedded Mouse breast carcinoma tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse breast carcinoma. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling Ki67 with ab279653 at 1/1000 dilution. The section was incubated with ab279653 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Counterstained with DAPI. Secondary antibody only control : Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution1) for 40 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat colon. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Ki67 with ab279653 at 1/1000 dilution. The section was incubated with ab279653 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Counterstained with DAPI. Secondary antibody only control : Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution1) for 40 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [B56] (AB279653)

Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling Ki67 with ab279653 at 1/1000 dilution. The section was incubated with ab279653 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Counterstained with DAPI. Secondary antibody only control : Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution1) for 40 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [B56] (AB279653)

ab279653 staining Ki67 in wild-type Hap1 cells, with negative expression in Ki67 knockout Hap1 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab279653 at 5 μg/ml and ab6046, Rabbit polyclonal to beta Tubulin at 1 μg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

This product also work with 4% formaldehyde (10 min) fixation under the same testing conditions.