Anti-NR2F2 antibody [EPR18443] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling NR2F2 with ab211777 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mesenchymal cells of human testis tissue (PMID : 11026559) 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) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 (human epithelial cell line from embryonic kidney) cells labeling NR2F2 with ab211777 at 1/100 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 on HEK293 cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab211777 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) secondary at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

Immunohistochemical analysis of paraffin-embedded human fetal spleen tissue labeling NR2F2 with ab211777 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mesenchymal cells of human fetal spleen tissue 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) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

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-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling NR2F2 with ab211777 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mesenchymal cells of human tonsil tissue (PMID : 11026559) 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) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

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-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling NR2F2 with ab211777 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on tumor cells of human breast cancer tissue 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) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

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-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling NR2F2 with ab211777 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on tumor cells of human colon cancer tissue 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) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

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-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling NR2F2 with ab211777 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mesenchymal cells of mouse liver tissue 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) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

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-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

Immunohistochemical analysis of paraffin-embedded rat lung tissue labeling NR2F2 with ab211777 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mesenchymal cells of rat lung tissue 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) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling NR2F2 with ab211777 at 1/100 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 on NIH/3T3 cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab211777 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150120) secondary at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

• WB

Lab

Western blot - Anti-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

This data was developed using the same antibody clone in a different buffer formulation (ab211777).

Lanes 1- 2 : Merged signal (red and green). Green - ab211777 observed at 45 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab211777 was shown to react with NR2F2 in wild-type HCT116 cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab266888 (CRISPR/Cas9 edited cell lysate ab257186) lane below 45kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NR2F2 CRISPR/Cas9 edited HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab211777 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NR2F2 antibody [EPR18443] (<a href='/en-us/products/primary-antibodies/nr2f2-antibody-epr18443-ab211777'>ab211777</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

NR2F2 CRISPR/Cas9 edited HCT116 cell lysate at 20 µg

Lane 2:

Western blot - Human NR2F2 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-nr2f2-knockout-hct116-cell-line-ab266888'>ab266888</a>)

Predicted band size: 46 kDa

Observed band size: 45 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

This data was created using ab211777, the same clone in a different buffer formulation.

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 K-562 (human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab211777 [EPR18443]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

This data was created using ab211777, the same clone in a different buffer formulation.

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 K-562 (human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab211777 [EPR18443]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

This data was created using ab211777, the same clone in a different buffer formulation.

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 K-562 (human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab211777 [EPR18443]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• IP

Supplier Data

Immunoprecipitation - Anti-NR2F2 antibody [EPR18443] - BSA and Azide free (AB240387)

NR2F2 was immunoprecipitated from 1 mg of MCF7 (human breast adenocarcinoma cell line) whole cell lysate with ab211777 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab211777 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : MCF7 whole cell lysate 10 μg (Input).
Lane 2 : ab211777 IP in MCF7 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab211777 in MCF7 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 second.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211777).

All lanes:

Immunoprecipitation - Anti-NR2F2 antibody [EPR18443] (<a href='/en-us/products/primary-antibodies/nr2f2-antibody-epr18443-ab211777'>ab211777</a>)

Predicted band size: 46 kDa

Observed band size: 46 kDa

true