Anti-Geminin antibody [EPR14637]

• ICC/IF

Supplier Data

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) (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 ab7291 (anti-Tubulin mouse mAb) at 1/1000 and 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 ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500.

• IHC-P

Supplier Data

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) (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 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.

• ICC/IF

AbReview59875****

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. ab150081 (1/200) was used as the secondary antibody.

This image is courtesy of an abreview by Kirk Mcmanus

• IHC-P

Supplier Data

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) (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 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.

• WB

Lab

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 ab264972 (knockout 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 (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 (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-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 (<a href='/en-us/products/cell-lines/human-gmnn-geminin-knockout-hela-cell-line-ab264972'>ab264972</a>)

Lane 3:

HepG2 lysate at 20 µg

Lane 4:

U-87 MG lysate at 20 µg

Predicted band size: 24 kDa

Observed band size: 32 kDa

false

• WB

Supplier Data

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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 24 kDa

false

Exposure time: 20s