Anti-Ki67 antibody [SP6] - BSA free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

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

ab16667 staining Ki67 in wild-type HAP1 cells (top panel) and Ki67 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol for 5 minutes, permeabilized 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 1 hour. The cells were then incubated with ab16667 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

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

Overlay histogram showing HAP1 wildtype (green line) and HAP1-MKI67 knockout cells (red line) stained with ab16667. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab16667, 1/1000) for 30 min at 22°C. The secondary antibody used wasGoat Anti-Rabbit IgG H&L (Alexa Fluor^®488) preadsorbed (ab150081) secondary antibodyat 1/2000 dilution for 30 min at 22°C. A Rabbit IgG isotype control antibody (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-MKI67 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. This image was generated from the hybridoma version.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

This data was developed using the same antibody clone in a different buffer formulation (ab16667). IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Human colon carcinoma tissue. The section incubated with ab16667 at 1/200 (0.145 μg/ml) dilution and ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880) was used as secondary antobody. Positive staining on human colon carcinoma. The section was incubated with ab16667 at 4°C overnight. The section was counterstained with haematoxylin. Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

This data was developed using the same antibody clone in a different buffer formulation (ab16667). Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Ki67 with ab16667 at 1/200 (0.145 µg/ml) followed by a ready to use LGoat Anti-Rabbit IgG H&L (HRP polymer) secondary antibody (ab214880). Low expression : positive staining only on proliferating cells (arrow) of human liver. The section was incubated with ab16667 at 4°C overnight and counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

This data was developed using the same antibody clone in a different buffer formulation (ab16667). Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Ki67 with ab16667 at 1/200 (0.145 µg/ml) followed by a ready to use LGoat Anti-Rabbit IgG H&L (HRP polymer) secondary antibody (ab214880). Low expression : positive staining only on proliferating cells (arrow) of human kidney. The section was incubated with ab16667 at 4°C overnight and counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

Tissue Microarrays stained for "Anti-Ki67 antibody [SP6]” using "ab16667" at 1/200 dilution (0.145 μg/ml) in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). The sections were incubated with ab16667 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Hematoxylin was used as the counter stain. This data was developed using the same antibody clone in a different buffer formulation (ab16667).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

This data was developed using the same antibody clone in a different buffer formulation (ab16667). Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Ki67 with ab16667 at 1/200 (0.145 µg/ml) followed by a ready to use LGoat Anti-Rabbit IgG H&L (HRP polymer) secondary antibody (ab214880). Low expression : positive staining only on proliferating cells (arrow) of mouse liver. The section was incubated with ab16667 at 4°C overnight and counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

This data was developed using the same antibody clone in a different buffer formulation (ab16667). Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Ki67 with ab16667 at 1/200 (0.145 µg/ml) followed by a ready to use LGoat Anti-Rabbit IgG H&L (HRP polymer) secondary antibody (ab214880). Low expression : positive staining only on proliferating cells (arrow) of mouse kidney. The section was incubated with ab16667 at 4°C overnight and counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880). Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0).

• IHC-P

AbReview23437****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

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

ab16667 staining Ki67 in rat small intestine tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using 10 mM citrate buffer pH 6.0. Samples were then blocked with 10% serum for 20 minutes at room temperature followed by incubation with the undiluted primary antibody for 30 minutes. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/2000 dilution.

This image was generated from the hybridoma version.

Image courtesy of Jing Ma by Abreview.

• WB

Lab

Western blot - Anti-Ki67 antibody [SP6] - BSA free (AB231172)

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. Loss of signal was observed when knockout sample 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

false