Anti-ETS1 antibody [EPR27353-81]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ETS1 antibody [EPR27353-81] (AB307672)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K562 (human chronic myelogenous leukemia lymphoblast, Left) / Daudi (human Burkitt's lymphoma lymphoblast, Right) cells labelling ETS1 with ab307672 at 1/50 dilution (1ug) (Red) (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 : K562 (PMID : 22107829)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETS1 antibody [EPR27353-81] (AB307672)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling ETS1 with ab307672 at 1/100 (5.3 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human tonsil. The section was incubated with ab307672 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ETS1 antibody [EPR27353-81] (AB307672)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell, Left) / Jurkat (human T cell leukemia T lymphocyte, Right) cells labelling ETS1 with ab307672 at 1/50 dilution (1ug) (Red) (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 : HepG2 (PMID : 30790340)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETS1 antibody [EPR27353-81] (AB307672)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscl tissue labeling ETS1 with ab307672 at 1/100 (5.3 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human skeletal muscle. The section was incubated with ab307672 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 was performed 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-ETS1 antibody [EPR27353-81] (AB307672)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling ETS1 with ab307672 at 1/100 (5.3 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on immune cells in human colon. The section was incubated with ab307672 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ETS1 antibody [EPR27353-81] (AB307672)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat (human T cell leukemia T lymphocyte) cells labelling ETS1 with ab307672 at 1/250 (2.12 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in Jurkat cell line, no staining was observed in HepG2 cell line.Negative control : HepG2 (PMID : 30790340).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). 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 2 ug/ml dilution.

• IP

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Immunoprecipitation - Anti-ETS1 antibody [EPR27353-81] (AB307672)

ETS1 was immunoprecipitated from 0.35 mg Daudi whole cell lysate with ab307672 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307672 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate Lane 2 : ab307672 IP in Daudi whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307672 in Daudi whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 58 seconds

All lanes:

Immunoprecipitation - Anti-ETS1 antibody [EPR27353-81] (ab307672) at 1/30 dilution

All lanes:

Daudi (human Burkitt's lymphoma lymphoblast) 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

false

Exposure time: 58s

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETS1 antibody [EPR27353-81] (AB307672)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling ETS1 with ab307672 at 1/100 (5.3 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse spleen (PMID : 17339335). The section was incubated with ab307672 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 was performed 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-ETS1 antibody [EPR27353-81] (AB307672)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling ETS1 with ab307672 at 1/100 (5.3 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat spleen. The section was incubated with ab307672 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-ETS1 antibody [EPR27353-81] (AB307672)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 169 seconds

All lanes:

Western blot - Anti-ETS1 antibody [EPR27353-81] (ab307672) at 1/1000 dilution

All lanes:

Human lymph node tissue lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Observed band size: 51 kDa

false

Exposure time: 169s

• WB

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Western blot - Anti-ETS1 antibody [EPR27353-81] (AB307672)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : HepG2 (PMID : 30790340) Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Exposure time : 125 seconds

All lanes:

Western blot - Anti-ETS1 antibody [EPR27353-81] (ab307672) at 1/1000 dilution

Lane 1:

Daudi (human Burkitt’s lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellar 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: 51 kDa

false

Exposure time: 125s

• WB

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Western blot - Anti-ETS1 antibody [EPR27353-81] (AB307672)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : skeletal muscle Exposure time : Lane 1 and 2 : 92 seconds, Lane 3 and 4 : 48 seconds

All lanes:

Western blot - Anti-ETS1 antibody [EPR27353-81] (ab307672) at 1/1000 dilution

Lane 1:

Mouse lymph node tissue lysate at 40 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 40 µg

Lane 3:

Rat lymph node tissue lysate at 40 µg

Lane 4:

Rat skeletal muscle tissue lysate at 40 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 1/20000 dilution

Observed band size: 51 kDa

false

Exposure time: 48s