Anti-NOL3 antibody [EPR25182-11]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NOL3 antibody [EPR25182-11] (AB288295)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Daudi (Human Burkitt's lymphoma lymphoblast, Left) / MCF7 (Human breast adenocarcinoma epithelial cell, Right) cells labelling NOL3 with ab288295 at 1/5000 dilution (0.01ug)(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.

Negative control : Daudi (PMID : HPA database).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NOL3 antibody [EPR25182-11] (AB288295)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 cells labelling NOL3 with ab288295 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in MCF7 cell line. Negative control : Daudi. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 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 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOL3 antibody [EPR25182-11] (AB288295)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling NOL3 with ab288295 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Cytoplasmic staining on human skeletal muscle. The section was incubated with ab288295 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).

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

• IP

Lab

Immunoprecipitation - Anti-NOL3 antibody [EPR25182-11] (AB288295)

NOL3 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab288295 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288295 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 : ab288295 IP in MCF7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288295 in MCF7 whole cell lysate

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

Exposure time : 32 seconds

All lanes:

Immunoprecipitation - Anti-NOL3 antibody [EPR25182-11] (ab288295) at 1/1000 dilution

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

Predicted band size: 24 kDa

Observed band size: 27 kDa

false

Exposure time: 32s

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NOL3 antibody [EPR25182-11] (AB288295)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a cells labelling NOL3 with ab288295 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in Neuro-2a cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 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 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOL3 antibody [EPR25182-11] (AB288295)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling NOL3 with ab288295 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond^®, Polymer Refine Detection) was used. Negative control : No staining on mouse liver. The section was incubated with ab288295 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOL3 antibody [EPR25182-11] (AB288295)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labelling NOL3 with ab288295 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond^®, Polymer Refine Detection). Positive staining on mouse cardiac muscle. The section was incubated with ab288295 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).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOL3 antibody [EPR25182-11] (AB288295)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labelling NOL3 with ab288295 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond^®, Polymer Refine Detection). Positive staining on rat cardiac muscle. The section was incubated with ab288295 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).

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

• IP

Lab

Immunoprecipitation - Anti-NOL3 antibody [EPR25182-11] (AB288295)

NOL3 was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab288295 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288295 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 10 ug

Lane 2 : ab288295 IP in Neuro-2a whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288295 in Neuro-2a whole cell lysate

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

Exposure time : 3 minutes

All lanes:

Immunoprecipitation - Anti-NOL3 antibody [EPR25182-11] (ab288295) at 1/1000 dilution

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

Predicted band size: 24 kDa

Observed band size: 27 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-NOL3 antibody [EPR25182-11] (AB288295)

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

The observed MW/expression profile is consistent with what has been described in the literature (PMID : 17142452).

Negative control : Daudi (PMID : HPA database)

Exposure time : 3 minutes

All lanes:

Western blot - Anti-NOL3 antibody [EPR25182-11] (ab288295) at 1/1000 dilution

Lane 1:

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

Lane 2:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 24 kDa

Observed band size: 27 kDa

false

• WB

Lab

Western blot - Anti-NOL3 antibody [EPR25182-11] (AB288295)

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

The observed MW/expression profile is consistent with what has been described in the literature (PMID : 15848180).

Exposure time : 15 seconds

All lanes:

Western blot - Anti-NOL3 antibody [EPR25182-11] (ab288295) at 1/1000 dilution

Lane 1:

Human heart tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 24 kDa

Observed band size: 27 kDa

false

• WB

Lab

Western blot - Anti-NOL3 antibody [EPR25182-11] (AB288295)

Blocking and diluting buffer : 5% NFDM/TBST.

The observed MW/expression profile is consistent with what has been described in the literature (PMID : 14645204, 20026055, 10590251).

Low expression : Mouse liver (PMID : 14645204)

Exposure time : 3 minutes

All lanes:

Western blot - Anti-NOL3 antibody [EPR25182-11] (ab288295) at 1/1000 dilution

Lane 1:

Mouse heart tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Rat heart tissue lysate at 20 µg

Lane 5:

Rat skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 24 kDa

Observed band size: 27 kDa

false