Anti-CIP2A antibody [EPR29632-7]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CIP2A antibody [EPR29632-7] (AB325239)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-2 OS (human bone osteosarcoma epithelial cell);SK-BR-3 (human breast adenocarcinoma epithelial cell) cells labelling CIP2A with ab325239 at 1/50 (9.98 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining and enriched in foci on condensed mitotic chromosomes in U-2 OS cells, with weak cytoplasmic staining SK-BR-3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : SK-BR-3(PMID : 24927563).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control1 : ab325239 at 1 : 50 dilution, followed by ab150120 at 1 : 1000 dilution
-ve control2 : ab7291 at 1 : 1000 dilution, followed by ab150081 at 1 : 1000 dilution

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CIP2A antibody [EPR29632-7] (AB325239)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling CIP2A with ab325239 at 1/100 (4.99 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon cancer (PMID : 32321509; PMID : 25896895). The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CIP2A antibody [EPR29632-7] (AB325239)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling CIP2A with ab325239 at 1/100 (4.99 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human breast cancer (PMID : 23306062). The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CIP2A antibody [EPR29632-7] (AB325239)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling CIP2A with ab325239 at 1/100 (4.99 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on human liver (PMID : 17632056). The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CIP2A antibody [EPR29632-7] (AB325239)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling CIP2A with ab325239 at 1/100 (4.99 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human testis (PMID : 17632056). The primary antibody was incubated 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

Lab

Immunoprecipitation - Anti-CIP2A antibody [EPR29632-7] (AB325239)

CIP2A was immunoprecipitated from 0.35 mg SiHa (human cervical epithelial cell) whole cell lysate with ab325239 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325239 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : SiHa (human cervical epithelial cell) whole cell lysate
Lane 2 : ab325239 IP in SiHa (human cervical epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325239 in SiHa whole cell lysate

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

Exposure time : 15 seconds.

All lanes:

Immunoprecipitation - Anti-CIP2A antibody [EPR29632-7] (ab325239) at 1/1000 dilution

Lane 1:

SiHa (human cervical epithelial cell) whole cell lysate at 10 µg

Lane 2:

ab325239 at 1/30 IP in SiHa (human cervical epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325239 in SiHa whole cell lysate

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

Observed band size: 90 kDa

false

Exposure time: 15s

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CIP2A antibody [EPR29632-7] (AB325239)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CIP2A with ab325239 at 1/50 (9.98 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining and enriched in foci on condensed mitotic chromosomes in NIH/3T3 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control1 : ab325239 at 1 : 50 dilution, followed by ab150120 at 1 : 1000 dilution
-ve control2 : ab7291 at 1 : 1000 dilution, followed by ab150081 at 1 : 1000 dilution

• IP

Lab

Immunoprecipitation - Anti-CIP2A antibody [EPR29632-7] (AB325239)

CIP2A was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab325239 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325239 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 2 : ab325239 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325239 in NIH/3T3 whole cell lysate

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

Exposure time : 8 seconds.

All lanes:

Immunoprecipitation - Anti-CIP2A antibody [EPR29632-7] (ab325239) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg

Lane 2:

ab325239 at 1/30 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325239 in NIH/3T3 whole cell lysate

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

Observed band size: 90 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-CIP2A antibody [EPR29632-7] (AB325239)

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

Low expression : liver, skeletal muscle(PMID : 17632056)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-CIP2A antibody [EPR29632-7] (ab325239) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human colon cancer tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Human breast cancer tissue lysate at 20 µg

Lane 5:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 90 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-CIP2A antibody [EPR29632-7] (AB325239)

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

Low expression : SK-BR-3(PMID : 24927563)

The identity of the bands below 75 kDa in lane 2 and lane 4 are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-CIP2A antibody [EPR29632-7] (ab325239) at 1/1000 dilution

Lane 1:

SiHa (human cervical epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

CT26.WT (mouse colon fibroblast) whole cell lysate at 20 µg

Lane 6:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 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 1/100000 dilution

Observed band size: 90 kDa,36 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-CIP2A antibody [EPR29632-7] (AB325239)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-CIP2A antibody [EPR29632-7] (ab325239) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting CIP2A whole cell lysate at 20 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 1/100000 dilution

Observed band size: 90 kDa,36 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-CIP2A antibody [EPR29632-7] (AB325239)

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

Low expression : liver, skeletal muscle(PMID : 17632056)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-CIP2A antibody [EPR29632-7] (ab325239) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 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 1/100000 dilution

Observed band size: 90 kDa,36 kDa

false

Exposure time: 180s