Anti-Cyclin B1 antibody [EPR17060]

11 product images

• 

  Western blot - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Lane 1: Wild type HAP1 whole cell lysate (20 μg)
  Lane 2: CCNB1 (KO) knockout HAP1 whole cell lysate (20 μg)
  Lane 3: HeLa whole cell lysate (20 μg)
  Lane 4: Hek293 whole cell lysate (20 μg)
  Lanes 1 - 4: Merged signal (red and green). Green - ab181593 observed at 55 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
  
  ab181593 was shown to recognize CCNB1 when CCNB1 knockout samples were used, along with additional cross-reactive bands. Wild-type and CCNB1 (KO) knockout samples were subjected to SDS-PAGE. ab181593 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
  
  
  

  All lanes: Western blot - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Predicted band size: 48 kDa

• 

  Flow Cytometry (Intracellular) - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Cyclin B1 with ab181593 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.
  

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Cyclin B1 with ab181593 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).

  Confocal image showing cytoplasm and weak nuclear staining on HeLa cell line.

  The nuclear counter stain is DAPI (blue).

  Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
  The negative controls are as follows:
  1. ab181593 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
  2. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (Mouse myoblast cell line) cells labeling Cyclin B1 with ab181593 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).

  Confocal image showing cytoplasm and nuclear staining on C2C12 cell line.

  The nuclear counter stain is DAPI (blue).

  Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
  The negative controls are as follows:
  1. ab181593 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
  2. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• 

  Western blot - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  5% NFDM/TBST: Blocking and diluting buffer.

  All lanes: Western blot - Anti-Cyclin B1 antibody [EPR17060] (ab181593) at 1/10000 dilution

  Lane 1: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

  Lane 2: Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution

  Predicted band size: 48 kDa

  Observed band size: 55 kDa

  Exposure time: 30s

• 

  Western blot - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  5% NFDM/TBST: Blocking and diluting buffer.

  All lanes: Western blot - Anti-Cyclin B1 antibody [EPR17060] (ab181593) at 1/2000 dilution

  Lane 1: C2C12 (Mouse myoblast cell line) whole cell lysate at 10 µg

  Lane 2: NIH 3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution

  Predicted band size: 48 kDa

  Observed band size: 55 kDa

  Exposure time: 30s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) was used as secondary at 1/500 dilution. Cytoplasm staining on the germinal center of Human tonsil is observed. Counter stained with Hematoxylin.
  

  Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Immunohistochemical analysis of paraffin-embedded Human lung squamous cell carcinomal tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) was used as secondary at 1/500 dilution. Cytoplasm staining on cancer cells of Human lung squamous cell carcinoma is observed. Counter stained with Hematoxylin.
  

  Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) was used as secondary at 1/500 dilution. Cytoplasm staining on epithelial cells of mouse colon is observed. Counter stained with Hematoxylin.
  

  Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) was used as secondary at 1/500 dilution. Negative staining on Human brain tissue. Counter stained with Hematoxylin.
  

  Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [EPR17060] (ab181593)

  Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Cyclin B1 using ab181593 at 1/500 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) was used as secondary at 1/500 dilution. Negative staining on Mouse liver tissue. Counter stained with Hematoxylin.
  

  Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.