Anti-Riz1 antibody [EPR25164-34]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Riz1 antibody [EPR25164-34] (AB305105)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling Riz1 with ab305105 at 1/500 dilution (0.1ug) (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). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Riz1 antibody [EPR25164-34] (AB305105)

Immunohistochemical analysis of paraffin-embedded Human lung tissue lABeling Riz1 with ab305105 at 1/16000 (0.032 ug/ml) followed by a ready to use Goat Anti-RABbit IgG H&L (HRP polymer). Nuclear staining on human lung. The section was incubated with ab305105 at 4ЎгC overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-RABbit IgG H&L (HRP polymer). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Riz1 antibody [EPR25164-34] (AB305105)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeABilized MCF7 (human breast adenocarcinoma epithelial cellЈ© cells lABelling Riz1 with ab305105 at 1/100 (5.09 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 MCF7 cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. 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

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Immunoprecipitation - Anti-Riz1 antibody [EPR25164-34] (AB305105)

Riz1 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab305105 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305105 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug
Lane 2 : ab305105 IP in MCF7 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305105 in MCF7 whole cell lysate

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

Exposure time : 8 seconds

Lysate was freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Immunoprecipitation - Anti-Riz1 antibody [EPR25164-34] (ab305105) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 μg

Lane 2:

MCF7 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: 260 kDa

false

Exposure time: 8s

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Riz1 antibody [EPR25164-34] (AB305105)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labelling Riz1 with ab305105 at 1/500 dilution (0.1ug) (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). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Riz1 antibody [EPR25164-34] (AB305105)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeABilized F9 (mouse embryonal carcinoma epithelial cell) cells lABelling Riz1 with ab305105 at 1/100 (5.09 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 F9 cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Riz1 antibody [EPR25164-34] (AB305105)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue lABeling Riz1 with ab305105 at 1/16000 (0.032 ug/ml) followed by a ready to use Goat Anti-RABbit IgG H&L (HRP polymer). Nuclear staining on mouse lung. The section was incubated with ab305105 at 4ЎгC overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-RABbit IgG H&L (HRP polymer). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• WB

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Western blot - Anti-Riz1 antibody [EPR25164-34] (AB305105)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTThe identity of the lower MW band at approximately 100 kDa is unknown. Exposure time : 10 seconds

All lanes:

Western blot - Anti-Riz1 antibody [EPR25164-34] (ab305105) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control, whole cell lysate 10 μg

Lane 2:

HeLa transfected with siRNA specifically targeti PR domain zinc fier protein 2, whole cell lysate 10 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: 260 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Riz1 antibody [EPR25164-34] (AB305105)

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

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The molecular weight observed is consistent with what has been described in the literature (PMID : 33585202).

Exposure time :

Lane 1 : 26 seconds

Lane 2 : 70 seconds

All lanes:

Western blot - Anti-Riz1 antibody [EPR25164-34] (ab305105) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

F9 (mouse embryonal carcinoma 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: 260 kDa

true