Anti-Geminin antibody [EPR14637]

6 product images

• 

  Western blot - Anti-Geminin antibody [EPR14637] (ab195047)

  ab195047 Anti-Geminin antibody [EPR14637] was shown to specifically react with Geminin DNA Replication Inhibitor in HeLa wild-type cells ( ab255448). Loss of signal was observed when knockout cell line Human GMNN (Geminin) knockout HeLa cell line ab264972 (knockout cell lysate Human GMNN (Geminin) knockout HeLa cell lysate ab257450) was used. Wild-type and Geminin DNA Replication Inhibitor knockout samples were subjected to SDS-PAGE. ab195047 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4°C at 1 in 500 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.

  All lanes: Western blot - Anti-Geminin antibody [EPR14637] (ab195047) at 1/500 dilution

  Lane 1: Wild-type HeLa lysate at 20 µg

  Lane 2: Geminin DNA Replication Inhibitor knockout HeLa lysate at 20 µg

  Lane 2: Western blot - Human GMNN (Geminin) knockout HeLa cell line (Human GMNN (Geminin) knockout HeLa cell line ab264972)

  Lane 3: HepG2 lysate at 20 µg

  Lane 4: U-87 MG lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 24 kDa

  Observed band size: 32 kDa

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Geminin antibody [EPR14637] (ab195047)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF-7 (Human breast adenocarcinoma cell line) cells labeling Geminin with ab195047 at 1/100 followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500(green). Confocal image showing nuclear staining on MCF-7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 (red).

  The negative controls are as follows:-
  -ve control 1 - ab195047 at 1/100 followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
  -ve control 2. - Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Geminin antibody [EPR14637] (ab195047)

  Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Geminin with ab195047 at 1/500 followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500. Nucleus staining on Human tonsil tissue is observed. Counter stained with Hematoxylin.
  

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

  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-Geminin antibody [EPR14637] (ab195047)

  Immunohistochemical analysis of paraffin-embedded Human triple-negative breast cancer tissue labeling Geminin with ab195047 at 1/500 followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500. Nucleus staining on Human triple-negative breast cancer tissue is observed. Counter stained with Hematoxylin.
  Negative control: Used PBS instead of primary antibody and Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 as secondary antibody.
  

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

• 

  This image is courtesy of a customer review by Kirk Mcmanus

  Immunocytochemistry/ Immunofluorescence - Anti-Geminin antibody [EPR14637] (ab195047)

  ab195047 staining Geminin in the HeLa cell line from Human cervix by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 Samples were incubated with primary antibody (1/50 in PBS ) for 1 hour at 22°C. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 (1/200) was used as the secondary antibody.

• 

  Western blot - Anti-Geminin antibody [EPR14637] (ab195047)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  Geminin is regulated during apoptosis by site-specific cleavage, resulting in the generation of two lower molecular weight cleavage products (21,462 ±2.7 Da and 25,082 ±2 Da). (PMID: 17261582)

  All lanes: Western blot - Anti-Geminin antibody [EPR14637] (ab195047) at 1/1000 dilution

  Lane 1: Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 15 µg

  Lane 2: HeLa treated with 2mM thymidine for 16 hours whole cell lysate at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 24 kDa

  Exposure time: 20s