Anti-PUS7 antibody [EPR25172-31]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PUS7 antibody [EPR25172-31] (AB289857)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methonol-permeabilised NIH/3T3 (Mouse embryonic fibroblast cell line) cells labelling PUS7 with ab289857 at 1/500 dilution (Red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG H&L (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-PUS7 antibody [EPR25172-31] (AB289857)

Immunohistochemical analysis of paraffin-embedded rat colon tissue lysate labeling PUS7 at 1/1000 (1.374μg/ml), followed by LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on rat colon. The section was incubated with ab289857 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counter stained with Hematoxylin.

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

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PUS7 antibody [EPR25172-31] (AB289857)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue lysate labeling PUS7 at 1/1000 (1.374μg/ml), followed by LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on mouse colon. The section was incubated with ab289857 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with Hematoxylin.

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

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PUS7 antibody [EPR25172-31] (AB289857)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labeling PUS7 with ab289857 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line. The nuclear counterstain is DAPI (Blue). Tubulin is labeled using ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution.

• WB

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Western blot - Anti-PUS7 antibody [EPR25172-31] (AB289857)

5% NFDM/TBST was used as blocking and diluting buffer.

Lysates were prepared from fresh material and used for Western blotting immediately to minimize protein degradation.
We observe an unknown band at around 37kDa.

All lanes:

Western blot - Anti-PUS7 antibody [EPR25172-31] (ab289857) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

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

Lane 4:

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

Lane 5:

HCT116 (human colorectal 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

Predicted band size: 75 kDa

Observed band size: 34 kDa,75 kDa

false

Exposure time: 15s

• WB

Unknown

Western blot - Anti-PUS7 antibody [EPR25172-31] (AB289857)

5% NFDM/TBST was used as blocking and diluting buffer.

Exposure times :

Lane 1 : 81 seconds
Lanes 2-3 : 180 seconds

All lanes:

Western blot - Anti-PUS7 antibody [EPR25172-31] (ab289857) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Rat pancreas tissue lysate at 20 µg

Lane 3:

Rat liver 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

Predicted band size: 75 kDa

Observed band size: 75 kDa

false