Anti-Geminin antibody [EPR23754-87]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Geminin antibody [EPR23754-87] (AB246509)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Geminin with ab246509 at 1/100 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human tonsil (PMID : 12107111). The section was incubated with ab246509 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Geminin antibody [EPR23754-87] (AB246509)

Intracellular flow cytometric analysis of 80% methanol fixed, 0.1% Tween-20 permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Geminin with ab246509 at 1/400 dilution (0.1μg) (Right panel) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left panel). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Cells were pre-treated with 20ug/ml RNaseA for 30 mins to minimize the binding between PI and RNA. Then intracellular stained with ab246509 and PI. Increase expression in S phase and G2/M phase was reported in literature (PMID : 28179914).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Geminin antibody [EPR23754-87] (AB246509)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HEK-293 (Human embryonic kidney epithelial cell) cells labelling Geminin with ab246509 at 1/400 dilution (0.1μg) (Right panel) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left panel). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Geminin antibody [EPR23754-87] (AB246509)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Geminin with ab246509 at 1/500 followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse testis (PMID : 22898305, PMID : 28690291). The section was incubated with ab246509 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Geminin antibody [EPR23754-87] (AB246509)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized MEF (Mouse embryonic fibroblast (immortalized)) cells labelling Geminin with ab246509 at 1/400 dilution (0.1μg) (Right panel) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left panel). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Geminin antibody [EPR23754-87] (AB246509)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Geminin with ab246509 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat testis (PMID : 22898305, PMID : 28690291). The section was incubated with ab246509 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Geminin antibody [EPR23754-87] (AB246509)

Immunohistochemical analysis of paraffin-embedded mouse breast carcinoma tissue labeling Geminin with ab246509 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse breast carcinoma (PMID : 12107111). The section was incubated with ab246509 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Unknown

Immunoprecipitation - Anti-Geminin antibody [EPR23754-87] (AB246509)

Geminin was immunoprecipitated from 0.35 mg mouse testis tissue lysate with ab246509 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246509 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse testis tissue lysate 10μg

Lane 2 : ab246509 IP in mouse testis tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab246509 in mouse testis tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-Geminin antibody [EPR23754-87] (ab246509)

Predicted band size: 24 kDa

Observed band size: 26 kDa

false

• WB

Lab

Western blot - Anti-Geminin antibody [EPR23754-87] (AB246509)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18798731).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-Geminin antibody [EPR23754-87] (ab246509) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 24 kDa

Observed band size: 26 kDa

false

• WB

Lab

Western blot - Anti-Geminin antibody [EPR23754-87] (AB246509)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18798731).

Exposure times. Lane 1, 4 : 15 seconds; Lane 2, 3 : 37 seconds; Lane 5 : 3 minutes.

All lanes:

Western blot - Anti-Geminin antibody [EPR23754-87] (ab246509) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 4:

Rat testis tissue lysate at 20 µg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 24 kDa

Observed band size: 26 kDa

false