Anti-Ki67 antibody [SP6] - BSA and Azide free

15 product images

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  Ki67 immunohistochemistry staining of human tonsil using rabbit anti-Ki67 antibody
  
  This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).
  Immunohistochemical analysis of formalin-fixed paraffin-embedded human tonsil sections labeling Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/200 dilution (0.156 μg/mL).

  Image A:
  Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer)
  Human tonsil tissue incubated with Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C.
  Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Citrate Buffer) ab93678 (citrate buffer pH 6.0).
  Nuclear staining on human tonsil. The section was incubated with Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C.

  Image B:
  Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection)
  Human tonsil tissue incubated with Anti-Ki67 antibody [SP6] ab16667 on a Leica Biosystems BOND^® RX instrument.
  Heat mediated antigen retrieval with Citrate buffer (pH 6.0 epitope retrieval solution2) for 20 mins.

• 

  Western blot - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).
  

  Lanes 1-3: Merged signal (red and green). Green - Anti-Ki67 antibody [SP6] ab16667 observed at 359 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007 observed at 125 kDa.

  Anti-Ki67 antibody [SP6] ab16667 was shown to react with Ki67 in wild-type HeLa. Loss of signal was observed when knockout sample Human MKI67 (Ki67) knockout HeLa cell lysate ab263762 was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. Anti-Ki67 antibody [SP6] ab16667 and Anti-Vinculin antibody VIN-54] (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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.

  Lanes 1 - 3: Western blot - Anti-Ki67 antibody [SP6] (Anti-Ki67 antibody [SP6] ab16667) at 1/100 dilution

  Lanes 1 - 3: Western blot - Anti-Ki67 antibody [SP6] - BSA free (Anti-Ki67 antibody [SP6] - BSA free ab231172) at 1/100 dilution

  Lanes 1 - 3: Western blot - Anti-Ki67 antibody [SP6], prediluted (Anti-Ki67 antibody [SP6], prediluted ab21700) at 1/100 dilution

  Lane 1: Ramos cell lysate at 20 µg

  Lane 2: Wild-type HeLa cell lysate at 20 µg

  Lane 3: MKI67 knockout HeLa cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 358 kDa

  Observed band size: 124 kDa, 359 kDa

  Lanes 1 - 3: Merged signal (red and green). Green - Anti-Ki67 antibody [SP6] ab16667 observed at 359 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007 observed at 125 kDa.
  

  Anti-Ki67 antibody [SP6] ab16667 was shown to react with Ki67 in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MKI67 (Ki67) knockout HeLa cell line ab255407 (knockout cell lysate Human MKI67 (Ki67) knockout HeLa cell lysate ab263762) was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. Anti-Ki67 antibody [SP6] ab16667 and Anti-Vinculin antibody [VIN-54] (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).
  

  IHC image of Anti-Ki67 antibody [SP6] ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Rat spleen tissue. The section was pre-treated using heat mediated antigen retrieval with (citrate buffer, pH 6.0). The section was then incubated with Anti-Ki67 antibody [SP6] ab16667, 1/200 (0.156 μg/mL) dilution and detected using ready to use Goat Anti-Rabbit IgG H&L (HRP) antibody. Nuclear staining on rat spleen. The section was incubated with Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  Ki67 immunofluorescence staining of HeLa cells using rabbit anti-Ki67 antibody
  
  This data was developed using Anti-Ki67 antibody [SP6] ab16667, the same antibody clone in a different buffer formulation.
  Immunofluorescent analysis of 100% methanol-fixed None permeabilized HeLa cells labelling Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1000 dilution (2 µg/mL).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).
  

  IHC image of Anti-Ki67 antibody [SP6] ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Human tonsil tissue. The section was pre-treated using heat mediated antigen retrieval with Antigen Retrieval Buffer (100X Citrate Buffer) ab93678 (citrate buffer, pH 6.0). The section was then incubated with Anti-Ki67 antibody [SP6] ab16667, 1/200 (0.156 μg/ml) dilution and detected using ready to use Goat Anti-Rabbit IgG H&L (HRP) antibody. Nuclear staining on human tonsil is observed. The section was incubated with Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  This data was developed using Anti-Ki67 antibody [SP6] ab16667, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 100% methanol-fixed, None permeabilized parental HAP1 cells labelling Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).

• 

  Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  Ki67 flow cytometry staining of HAP1 cells using rabbit anti-Ki67 antibody
  
  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 Anti-Ki67 antibody [SP6] ab16667 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  This data was developed using Anti-Ki67 antibody [SP6] ab16667, the same antibody clone in a different buffer formulation.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).
  

  IHC image of Anti-Ki67 antibody [SP6] ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Mouse spleen tissue. The section was pre-treated using heat mediated antigen retrieval with Antigen Retrieval Buffer (100X Citrate Buffer) ab93678 (citrate buffer, pH 6.0). The section was then incubated with Anti-Ki67 antibody [SP6] ab16667, 1/200 (0.156 μg/mL) dilution and detected using a ready to use Goat Anti-Rabbit IgG H&L (HRP) antibody. Nuclear staining on mouse spleen.The section was incubated with Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).
  

  Anti-Ki67 antibody [SP6] 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 Anti-Ki67 antibody [SP6] ab16667 at 1/250 dilution and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 (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.

  This image was generated from the hybridoma version.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).
  

  Anti-Ki67 antibody [SP6] ab16667 staining Ki67 in rat oesophagus by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then blocked with 1% BSA for 10 minutes at 21°C followed by incubation with the primary antibody for 30 minutes at 1/100. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

  This image was generated from the hybridoma version.

• 

  Multiplex immunohistochemistry - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  This data was developed using Anti-Ki67 antibody [SP6] ab16667, the same antibody clone in a different buffer formulation.
  
  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with Anti-PD1 antibody [EPR23119-111] ab243644 at 1/500 dilution (1.02 µg/mL) (D), Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/200 dilution (0.15 μg/ml) (B) and PD-L1 with Anti-PD-L1 antibody [SP142] - C-terminal ab228462 at 1/100 dilution (0.52 μg/ml) (C). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
  
  Panel A: merged staining of anti-Ki67 (magenta; Opal™690), anti-PD-L1 (red; Opal™570) and anti-PD1 (green; Opal™520) on human tonsil.
  Panel B: anti-Ki67 stained on nucleus of proliferating cells.
  Panel C: anti-PD-L1 stained on membrane of cells involved in T cell inhibition.
  Panel D: anti-PD1 stained on antigen-stimulated T cells.
  
  The section was incubated in three rounds of staining: in the order of Anti-Ki67 antibody [SP6] ab16667 for 10 mins, Anti-PD1 antibody [EPR23119-111] ab243644 for 30 mins and Anti-PD-L1 antibody [SP142] - C-terminal ab228462 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• 

  Western blot - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  This data was developed using the same antibody clone in a different buffer formulation (abAB16667).

  Western blot: Anti-MKI67 antibody [SP6] (Anti-Ki67 antibody [SP6] ab16667) staining at 1/500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Ki67 antibody [SP6] ab16667 was shown to bind specifically to MKI67. A band was observed at 359 kDa in wild-type A549 cell lysates with no signal observed at this size in MKI67 knockout cell line. To generate this image, wild-type and MKI67 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Ki67 antibody [SP6] (Anti-Ki67 antibody [SP6] ab16667) at 1/500 dilution

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: MKI67 knockout A549 cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 359 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil Ki67 with ab197547 at a concentration of 5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab197547 anti Ki67 antibody was incubated at 37oC for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  Tissue Microarrays stained for "Anti-Ki67 antibody [SP6]” using "Anti-Ki67 antibody [SP6] 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 Antigen Retrieval Buffer (100X Citrate Buffer) ab93678 (citrate buffer, pH 6.0). The sections were incubated with Anti-Ki67 antibody [SP6] ab16667 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (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 (Anti-Ki67 antibody [SP6] ab16667).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

  Ki67 immunohistochemistry staining of human tonsil using rabbit anti-Ki67 antibody
  
  

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).

  Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/500 dilution. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). Anti-Ki67 antibody [SP6] ab16667 anti Ki67 antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.