Anti-NOP2 antibody [EPR24888-92]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOP2 antibody [EPR24888-92] (AB271075)

Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma tissue labelling NOP2 with ab271075 at 1/200 dilution (2.835 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on human lung adenocarcinoma (PMID : 3422591). The section was incubated with ab271075 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) was used.

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOP2 antibody [EPR24888-92] (AB271075)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labelling NOP2 with ab271075 at 1/200 dilution (2.835 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nucleolar staining on human cerebrum (PMID : 25481415). The section was incubated with ab271075 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) was used.

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NOP2 antibody [EPR24888-92] (AB271075)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HCT 116 (human colorectal carcinoma cell line) cells labelling NOP2 with ab271075 at 1/500 dilution (1.134 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/ml / Green). Confocal image showing nucleolar staining in HCT 116 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 μg/ml / Red). 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 (2 μg/ml) dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-NOP2 antibody [EPR24888-92] (AB271075)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cells labelling NOP2 with ab271075 at 1/50 dilution (1μg / 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, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-NOP2 antibody [EPR24888-92] (AB271075)

NOP2 was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate with ab271075 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271075 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 293T (human embryonic kidney epithelial cell) whole cell lysate 10 μg

Lane 2 : ab271075 IP in 293T whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271075 in 293T whole cell lysate

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

Exposure time : 24 seconds

All lanes:

Immunoprecipitation - Anti-NOP2 antibody [EPR24888-92] (ab271075)

Predicted band size: 89 kDa

Observed band size: 110 kDa,90 kDa

false

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NOP2 antibody [EPR24888-92] (AB271075)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labelling NOP2 with ab271075 at 1/500 dilution (1.134 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/ml / Green). Confocal image showing nucleolar staining in Neuro-2a cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 μg/ml / Red). 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 dilution (2 μg/ml).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOP2 antibody [EPR24888-92] (AB271075)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labelling NOP2 with ab271075 at 1/500 dilution (1.134 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nucleolar staining on mouse cerebrum. The section was incubated with ab271075 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) was used.

Heat mediated antigen retrieval 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-NOP2 antibody [EPR24888-92] (AB271075)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labelling NOP2 with ab271075 at 1/500 dilution (1.134 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nucleolar staining on rat cerebrum. The section was incubated with ab271075 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) was used.

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

• WB

Lab

Western blot - Anti-NOP2 antibody [EPR24888-92] (AB271075)

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

Lysates were made freshly and used in WB test immediately to minimize protein degradation.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 25043274).

The multiple reactive bands would represent different NOP2 isoforms.

Exposure time : 48 seconds

All lanes:

Western blot - Anti-NOP2 antibody [EPR24888-92] (ab271075) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

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 1/50000 dilution

Predicted band size: 89 kDa

Observed band size: 110 kDa,90 kDa

false