重组Anti-PABPN1抗体[EP3000Y]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling PABPN1 with unpurified ab75855 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
Control : PBS only.
Nuclear counter stain : DAPI.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling PABPN1 with purified ab75855 at 1/40 dilution (10 ug/ml) (red). Cells were fixed with 80% Methanol and permeabilized with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluorr^®488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue sections labeling PABPN1 with Purified ab75855 at 1 : 1000 dilution (0.41 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Overlay histogram showing MCF-7 cells stained with unpurified ab75855 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75855, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• IP

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Immunoprecipitation - Anti-PABPN1 antibody [EP3000Y] (AB75855)

ab75855 (purified) at 1 : 30 dilution (5ug) immunoprecipitating PABPN1 in MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate.
Lane 1 (input) : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+) : ab75855 & MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab75855 in MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 10,000 dilution.

All lanes:

Immunoprecipitation - Anti-PABPN1 antibody [EP3000Y] (ab75855)

Predicted band size: 33 kDa

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• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling PABPN1 with Purified ab75855 at 1 : 1000 dilution (0.41 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling PABPN1 with Purified ab75855 at 1 : 1000 dilution (0.41 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• WB

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Western blot - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Lanes 1 - 2:

Western blot - Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/200000 dilution

Lane 3:

Western blot - Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/1000000 dilution

Lane 1:

Raw264.7 cell lysate at 10 µg

Lane 2:

MCF-7 cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/1000 dilution

Predicted band size: 33 kDa

Observed band size: 49 kDa

false

• WB

Lab

Western blot - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/2000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) whole cell lysates at 15 µg

Lane 2:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 33 kDa

Observed band size: 49 kDa

false

• WB

Lab

Western blot - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/2000 dilution

All lanes:

Rat brain lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 33 kDa

false

• WB

Lab

Western blot - Anti-PABPN1 antibody [EP3000Y] (AB75855)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/2000 dilution

Lane 1:

RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 15 µg

Lane 2:

Mouse spleen lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 33 kDa

false