Anti-ZCWCC1 antibody [EPR28655-56]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZCWCC1 antibody [EPR28655-56] (AB315903)

Immunohistochemical analysis of paraffin-embedded Human stomach cancer tissue labeling ZCWCC1 with ab315903 at 1/100 (5.03 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Nuclear staining on human stomach cancer (PMID : 26098774). The section was incubated with ab315903 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 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-ZCWCC1 antibody [EPR28655-56] (AB315903)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling ZCWCC1 with ab315903 at 1/100 (5.03 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Nuclear staining on human breast (PMID : 29555576). The section was incubated with ab315903 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 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-ZCWCC1 antibody [EPR28655-56] (AB315903)

Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling ZCWCC1 with ab315903 at 1/100 (5.03 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Nuclear staining on human stomach (PMID : 26098774). The section was incubated with ab315903 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ZCWCC1 antibody [EPR28655-56] (AB315903)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling ZCWCC1 with ab315903 at 1/50 (10.06 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing nuclear staining in HeLa cells.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ZCWCC1 antibody [EPR28655-56] (AB315903)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling ZCWCC1 with ab315903 at 1/500 dilution (0.1 ug)/Red (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.

• IP

Supplier Data

Immunoprecipitation - Anti-ZCWCC1 antibody [EPR28655-56] (AB315903)

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

ZCWCC1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab315903 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315903 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab315903 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315903 in HeLa whole cell lysate

Lysates were freshly made and used for IP immediately to minimize protein degradation.

All lanes:

Immunoprecipitation - Anti-ZCWCC1 antibody [EPR28655-56] (ab315903) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

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 1/1000 dilution

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ZCWCC1 antibody [EPR28655-56] (AB315903)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling ZCWCC1 with ab315903 at 1/50 (10.06 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing nuclear staining in Neuro-2a cells.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-ZCWCC1 antibody [EPR28655-56] (AB315903)

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

ZCWCC1 was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab315903 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315903 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate
Lane 2 : ab315903 IP in Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315903 in Neuro-2a whole cell lysate

Lysates were freshly made and used for IP immediately to minimize protein degradation.

All lanes:

Immunoprecipitation - Anti-ZCWCC1 antibody [EPR28655-56] (ab315903) at 1/30 dilution

All lanes:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 10 µg

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 1/1000 dilution

false

• WB

Supplier Data

Western blot - Anti-ZCWCC1 antibody [EPR28655-56] (AB315903)

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

The bands beneath the target band around 65-85kDa are likely to be degraded target fragments.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

Exposure time : Lanes 1-5 : 10 seconds; Lane 6 : 37 seconds.

All lanes:

Western blot - Anti-ZCWCC1 antibody [EPR28655-56] (ab315903) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

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

Lane 4:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lanes 5 - 6:

T-47D (human ductal breast epithelial tumor 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 1/20000 dilution

Observed band size: 135 kDa

false

• WB

Supplier Data

Western blot - Anti-ZCWCC1 antibody [EPR28655-56] (AB315903)

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

The bands beneath the target band around 65-85kDa are likely to be degraded target fragments.

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

All lanes:

Western blot - Anti-ZCWCC1 antibody [EPR28655-56] (ab315903) 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 ZCWCC1 whole cell lysate at 20 µg

Lane 3:

T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (human breast 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 1/20000 dilution

Observed band size: 135 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-ZCWCC1 antibody [EPR28655-56] (AB315903)

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

The bands beneath the target band are likely to be degraded target fragments.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-ZCWCC1 antibody [EPR28655-56] (ab315903) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat testis tissue lysate at 20 µg

Lane 3:

Rat spleen tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Mouse testis 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 1/20000 dilution

Observed band size: 135 kDa

false

Exposure time: 180s