Anti-Cdc20 antibody [EPR28091-55]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil. The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on human cerebrum. The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human ovarian carcinoma. The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) treated with 100ng/mL nocodazole for 16 hours (Red) / Untreated HeLa (Dotted Red) cells labelling Cdc20 with ab317036 at 1/50 dilution (1 ug) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488 ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon (PMID : 26245990). The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Cdc20 was immunoprecipitated from 0.35 mg PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate with ab317036 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317036 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab317036 IP in PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317036 in PC-3 whole cell lysate

The band at ~37KDa may represent a truncated Cdc20 isoform (PMID : 37100900).

All lanes:

Immunoprecipitation - Anti-Cdc20 antibody [EPR28091-55] (ab317036) at 1/30 dilution

All lanes:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse colon. The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on rat cerebrum. The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on mouse cerebrum. The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Cdc20 with ab317036 at 1/50 dilution (1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488 ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat testis. The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat colon. The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Cdc20 with ab317036 at 1/100 (4.96 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse testis. The section was incubated with ab317036 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

The expression of Cdc20 is upregulated in response to nocodazole treatment. The band at ~37KDa may represent a truncated Cdc20 isoform (PMID : 37100900).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Cdc20 antibody [EPR28091-55] (ab317036) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

HeLa treated with 100ng/ml nocodazole for 16 hours whole cell lysate at 20 µg with 5% NFDM/TBST

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 1/100000 dilution

Observed band size: 51 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Low expression : DU145 (PMID : 27633058), LNCaP (PMID : 27633058).

In lanes 1-4, the lysates were stored at -80°C prior to Western Blotting. In lanes 5-8, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Cdc20 antibody [EPR28091-55] (ab317036) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 40 µg with 5% NFDM/TBST

Lane 2:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 40 µg with 5% NFDM/TBST

Lane 3:

DU145 (human prostate carcinoma epithelial cell) whole cell lysate at 40 µg with 5% NFDM/TBST

Lane 4:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 40 µg with 5% NFDM/TBST

Lane 5:

HeLa whole fresh cell lysate at 20 µg with 5% NFDM/TBST

Lane 6:

293T (human embryonic kidney epithelial cell) whole fresh cell lysate at 20 µg with 5% NFDM/TBST

Lane 7:

NIH/3T3 (mouse embryonic fibroblast) whole fresh cell lysate at 20 µg with 5% NFDM/TBST

Lane 8:

PC-12 (rat adrenal gland pheochromocytoma) whole fresh cell lysate at 20 µg with 5% NFDM/TBST

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 1/100000 dilution

Observed band size: 51 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Cdc20 antibody [EPR28091-55] (AB317036)

Low expression : kidney (PMID : 15280373), liver (PMID : 15280373), brain (PMID : 15280373).

In lanes 1-6, the lysates were stored at -80°C prior to Western Blotting. In lanes 7-11, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

The identity of the higher MW bands at approximately 74 kDa (in lanes 1-7) and 90 KDa (lane 8) are unknown.

Lanes 4-11 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjMSD1483ated (ab97051) at 1/100000 and lanes 1-3 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-6, 9-11 : 180 seconds, lanes 7-8 : 70 seconds.

All lanes:

Western blot - Anti-Cdc20 antibody [EPR28091-55] (ab317036) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 40 µg with 5% NFDM/TBST

Lane 2:

Human kidney tissue lysate at 40 µg with 5% NFDM/TBST

Lane 3:

Human liver tissue lysate at 40 µg with 5% NFDM/TBST

Lane 4:

Mouse testis tissue lysate at 40 µg with 5% NFDM/TBST

Lane 5:

Mouse thymus tissue lysate at 40 µg with 5% NFDM/TBST

Lane 6:

Mouse kidney tissue lysate at 40 µg with 5% NFDM/TBST

Lane 7:

Mouse testis tissue lysate at 20 µg with 5% NFDM/TBST

Lane 8:

Mouse brain tissue lysate at 20 µg with 5% NFDM/TBST

Lane 9:

Rat testis tissue lysate at 20 µg with 5% NFDM/TBST

Lane 10:

Rat brain tissue lysate at 20 µg with 5% NFDM/TBST

Lane 11:

Rat liver tissue lysate at 20 µg with 5% NFDM/TBST

Secondary

Lanes 1 - 3:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 4 - 11:

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 1/100000 dilution

Observed band size: 51 kDa,36 kDa

false