Anti-RUNX3 antibody [EPR20680]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20680] (AB224642)

Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling RUNX3 with ab224642 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on lymphoid cells of human stomach (PMID : 15514019; PMID : 21786422) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20680] (AB224642)

Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling RUNX3 with ab224642 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on lymphoid cells of human gastric cancer (PMID : 27566570). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX3 antibody [EPR20680] (AB224642)

Immunohistochemical analysis of paraffin-embedded human diffuse large B cell lymphoma tissue labeling RUNX3 with ab224642 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on human diffuse large B cell lymphoma (PMID : 27184221). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Unknown

Immunoprecipitation - Anti-RUNX3 antibody [EPR20680] (AB224642)

RUNX3 was immunoprecipitated from 0.35 mg of Raji (human Burkitt's lymphoma cell line) whole cell lysate with ab224642 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224642 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Raji whole cell lysate 10 μg (Input).

Lane 2 : ab224642 IP in Raji whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab224642 in Raji whole cell lysate.

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

Exposure time : 10 second.

The molecular weight observed is consistent with what has been described in the literature (PMID : 23700080).

All lanes:

Immunoprecipitation - Anti-RUNX3 antibody [EPR20680] (ab224642)

Predicted band size: 44 kDa

false

• WB

Supplier Data

Western blot - Anti-RUNX3 antibody [EPR20680] (AB224642)

Exposure time : Lane 1 : 3 minutes; Lane 2 : 15 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

This target detects both predicted isoforms 44 kDa and 46 kDa, consistent with what has been described in the literature (PMID : 23700080).

All lanes:

Western blot - Anti-RUNX3 antibody [EPR20680] (ab224642) at 1/1000 dilution

Lane 1:

Human thymus tissue lysate at 20 µg

Lane 2:

Human tonsil tissue 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

Predicted band size: 44 kDa

Observed band size: 44 kDa,46 kDa

true

• WB

Lab

Western blot - Anti-RUNX3 antibody [EPR20680] (AB224642)

Lanes 1 - 4 : Merged signal (red and green). Green - ab224642 observed at 44-46 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab224642 was shown to recognize RUNX3 in wild-type HAP1 cells as signal was lost at the expected MW in RUNX3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and RUNX3 knockout samples were subjected to SDS-PAGE. ab224642 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-RUNX3 antibody [EPR20680] (ab224642) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

RUNX3 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

Ramos whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Predicted band size: 44 kDa

false

• WB

Supplier Data

Western blot - Anti-RUNX3 antibody [EPR20680] (AB224642)

Blocking/dilution buffer : 5% NFDM/TBST.

Negative control : : MCF-7 (PMID : 21706051).
This target detects both predicted isoforms 44KDa and 46KDa, consistent with what has been described in the literature (PMID : 23700080).

All lanes:

Western blot - Anti-RUNX3 antibody [EPR20680] (ab224642) at 1/1000 dilution

Lane 1:

Raji (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 2:

Ramos (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma cell line) 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

Predicted band size: 44 kDa

Observed band size: 44 kDa,46 kDa

true

Exposure time: 10s