Recombinant Anti-ETS2 antibody [EPR22419] (ab219948)

Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling ETS2 with ab219948 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on mouse thymus (PMID: 3472202, PMID:26926107) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded rat thymus tissue labeling ETS2 with ab219948 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on rat thymus (PMID: 3472202, PMID:26926107) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling ETS2 with ab219948 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in RAW 264.7 cells treated with lipopolysaccharide (500 ng/ml) for 6 hours. The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
Secondary antibody contril: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling ETS2 with ab219948 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Negative control: No staining on rat cerebellum (PMID: 3472202) is observed. Counterstained with hematoxylin.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling ETS2 with ab219948 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Negative control: No staining on mouse cerebrum (PMID: 3472202) is observed. Counterstained with hematoxylin.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

ETS2 was immunoprecipitated from 0.35 mg K562 (human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate with ab219948 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219948 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: K562 whole cell lysate 10 µg (Input).
Lane 2: ab219948 IP in K562 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219948 in K562 whole cell lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.

The two immunoprecipitated bands represent non-phosphorylated and phosphorylated ETS2.

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lane 1: 26 seconds, Lane 2: 3 minutes.

Lane 1 was developed using a higher sensitivity ECL substrate.

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID: 15590657). The doublets represent ETS2 and its phosphorylated form.

This blot was developed using a higher sensitivity ECL substrate.