Anti-FOXP1 antibody [RM1113]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling FOXP1 with ab320745 at 1/1000 (0.509 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human tonsil. The section was incubated with ab320745 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling FOXP1 with ab320745 at 1/1000 (0.509 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression : weak staining on human skeletal muscle. The section was incubated with ab320745 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling FOXP1 with ab320745 at 1/50 (10.18 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 MCF7 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Negative control : HDLM-2 (PMID : 16200457). 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 (2.5 ug/ml) dilution (Magenta). 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.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-FOXP1 antibody [RM1113] (AB320745)

Chromatin was prepared from K-562 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10e7 cells and 8 µg of ab320745 [RM1113]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-FOXP1 antibody [RM1113] (AB320745)

Chromatin was prepared from K-562 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10e7 cells and 8 µg of ab320745 [RM1113]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-FOXP1 antibody [RM1113] (AB320745)

Chromatin was prepared from K-562 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10e7 cells and 8 µg of ab320745 [RM1113]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

Supplier Data

Immunoprecipitation - Anti-FOXP1 antibody [RM1113] (AB320745)

FOXP1 was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab320745 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab320745 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution. Lane 1 : K-562 whole cell lysate Lane 2 : ab320745 IP in K-562 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab320745 in K-562 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 58 seconds.

All lanes:

Immunoprecipitation - Anti-FOXP1 antibody [RM1113] (ab320745) at 1/30 dilution

All lanes:

K-562 (human chronic myelogenous leukemia 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling FOXP1 with ab320745 at 1/5000 (0.509 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression : weak staining on mouse skeletal muscle. The section was incubated with ab320745 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skeletal muscle (fresh frozen) tissue labeling FOXP1 with ab320745 at 1/50 (10.18 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Low expression : confocal image showing no staining on mouse skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab320745 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat skeletal muscle (fresh frozen) tissue labeling FOXP1 with ab320745 at 1/50 (10.18 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Low expression : confocal image showing no staining on rat skeletal muscle. The nuclear counterstain was DAPI (Blue). The section was incubated with ab320745 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling FOXP1 with ab320745 at 1/5000 (0.102 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse spleen. The section was incubated with ab320745 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling FOXP1 with ab320745 at 1/5000 (0.509 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Low expression : weak staining on rat skeletal muscle. The section was incubated with ab320745 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling FOXP1 with ab320745 at 1/5000 (0.102 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat spleen. The section was incubated with ab320745 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat spleen (fresh frozen) tissue labeling FOXP1 with ab320745 at 1/50 (10.18 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing positive staining on rat spleen. The nuclear counterstain was DAPI (Blue). The section was incubated with ab320745 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-FOXP1 antibody [RM1113] (AB320745)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen (fresh frozen) tissue labeling FOXP1 with ab320745 at 1/50 (10.18 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing positive staining on mouse spleen. The nuclear counterstain was DAPI (Blue). The section was incubated with ab320745 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-FOXP1 antibody [RM1113] (AB320745)

FOXP1 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab320745 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab320745 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution. Lane 1 : RAW 264.7 whole cell lysate Lane 2 : ab320745 IP in RAW 264.7 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab320745 in RAW 264.7 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 24 seconds. To minimize protein degradation, cells and tissues were lysed immediately after harvest and loaded onto membranes for IP.

All lanes:

Immunoprecipitation - Anti-FOXP1 antibody [RM1113] (ab320745) at 1/30 dilution

All lanes:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) 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: 24s

• WB

Supplier Data

Western blot - Anti-FOXP1 antibody [RM1113] (AB320745)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : HDLM-2. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 12692134, PMID : 24416450). The band below 37 kDa may be caused by degradation. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control  (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-FOXP1 antibody [RM1113] (ab320745) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

Ramos (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 20 µg

Lane 4:

WEHI-231 (mouse B cell lymphoma B lymphocyte ) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 50 kDa,60 kDa,70 kDa,95 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-FOXP1 antibody [RM1113] (AB320745)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 12692134, PMID : 24416450). The band below 37 kDa may be caused by degradation. The lysates are suggested freshly made and used for Western Blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-FOXP1 antibody [RM1113] (ab320745) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Secondary

All lanes:

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

Observed band size: 50 kDa,60 kDa,70 kDa,95 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-FOXP1 antibody [RM1113] (AB320745)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : skeletal muscle (PMID : 12692134). The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 12692134, PMID : 24416450). The band below 37 kDa may be caused by degradation. The lysates are suggested freshly made and used for Western Blotting immediately to minimize protein degradation (Lane 1 and Lane 7). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control  (ab181602) staining at 1/200000 dilution. Exposure time : Lane 1-6 : 48 seconds Lane 7 : 180 seconds

All lanes:

Western blot - Anti-FOXP1 antibody [RM1113] (ab320745) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell frozen lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

2.4G2 (mouse/rat hybridoma) whole cell lysate at 20 µg

Lane 5:

Rat spleen tissue lysate at 20 µg

Lane 6:

Rat skeletal muscle tissue lysate at 20 µg

Lane 7:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell fresh 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 1/20000 dilution

Observed band size: 50 kDa,60 kDa,70 kDa,95 kDa,36 kDa

false