Anti-Ki67 antibody [SP6], prediluted

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6], prediluted (AB21700)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized parental HAP1 (Wildtype controlHuman chronic myelogenous leukemianear-haploid cell line) cells labelling Ki67 with ab16667 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730)(Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488 ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using ab16667, the same antibody clone in a different buffer formulation.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (AB21700)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human thymus tissue, staining Ki67 with ab21700.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (AB21700)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human breast adenocarcinoma tissue, staining Ki67 with ab21700.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6], prediluted (AB21700)

This data was developed using ab16667, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 100% methanol-fixed None permeabilized HeLa cells labelling Ki67 with ab16667 at 1/1000 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing nucleolar staining in HeLa cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1000 dilution (2 µg/mL).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6], prediluted (AB21700)

This data was developed using ab16667, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 100% methanol-fixed, None permeabilized parental HAP1 cells labelling Ki67 with ab16667 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing nucleolar staining in parental HAP1cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (AB21700)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining Ki67 with ab21700.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (AB21700)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human endometrial adenocarcinoma tissue, staining Ki67 with ab21700.

• WB

Lab

Western blot - Anti-Ki67 antibody [SP6], prediluted (AB21700)

This data was developed using the same antibody clone in a different buffer formulation (ab16667).

Lanes 1 - 3 : Merged signal (red and green). Green - ab16667 observed at 359 kDa. Red - loading control, ab130007 observed at 125 kDa.

ab16667 was shown to react with Ki67 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255407 (knockout cell lysate ab263762) was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. ab16667 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 100 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Ki67 antibody [SP6] (<a href='/en-us/products/primary-antibodies/ki67-antibody-sp6-ab16667'>ab16667</a>) at 1/100 dilution

Lane 1:

Ramos cell lysate at 20 µg

Lane 2:

Wild-type HeLa cell lysate at 20 µg

Lane 3:

Western blot - Human MKI67 (Ki67) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-mki67-ki67-knockout-hela-cell-line-ab255407'>ab255407</a>) at 20 µg

Lane 3:

Western blot - Human MKI67 (Ki67) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-mki67-ki67-knockout-hela-cell-lysate-ab263762'>ab263762</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 358 kDa

Observed band size: 124 kDa,359 kDa

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