Anti-RUNX1 / AML1 antibody [RM1089]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX1 / AML1 antibody [RM1089] (AB315398)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling RUNX1 / AML1 with ab315398 at 1/500 (1.092 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on the human liver (PMID : 34376784). The section was incubated with ab315398 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-RUNX1 / AML1 antibody [RM1089] (AB315398)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling RUNX1 / AML1 with ab315398 at 1/500 (1.092 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on the human tonsil. The section was incubated with ab315398 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-RUNX1 / AML1 antibody [RM1089] (AB315398)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling RUNX1 / AML1 with ab315398 at 1/500 (1.092 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on the human breast carcinoma. The section was incubated with ab315398 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX1 / AML1 antibody [RM1089] (AB315398)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling RUNX1 / AML1 with ab315398 at 1/500 (1.092 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on the human breast. The section was incubated with ab315398 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-RUNX1 / AML1 antibody [RM1089] (AB315398)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labelling RUNX1 / AML1 with ab315398 at 1/50 (10.92 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, and HEK293 cell line showed nearly no staining.
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 1/500 (1ug/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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RUNX1 / AML1 antibody [RM1089] (AB315398)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK293 (human embryonic kidney epithelial cell, Left) / Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling RUNX1 / AML1 with ab315398 at 1/40000 dilution (0.00125 ug)/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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : HEK293.

• IP

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Immunoprecipitation - Anti-RUNX1 / AML1 antibody [RM1089] (AB315398)

RUNX1 / AML1 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate with ab315398 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315398 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-RUNX1 / AML1 antibody [RM1089] (ab315398) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate

Lane 2:

ab315398 at 1/30 IP in Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab315398 in Jurkat 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: 10s

• WB

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Western blot - Anti-RUNX1 / AML1 antibody [RM1089] (AB315398)

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

Low expression : HEK-293.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 17431130 and 29296779).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-RUNX1 / AML1 antibody [RM1089] (ab315398) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 2:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 3:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

THP-1 (human monocytic leukemia monocyte) 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: 27-55 kDa,15 kDa

false

Exposure time: 180s

• ChIC/CUT&RUN-seq

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ChIC/CUT&RUN sequencing - Anti-RUNX1 / AML1 antibody [RM1089] (AB315398)

ChIC/CUT&RUN sequencing 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 ab315398 [RM1089]. 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

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ChIC/CUT&RUN sequencing - Anti-RUNX1 / AML1 antibody [RM1089] (AB315398)

ChIC/CUT&RUN sequencing 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 ab315398 [RM1089]. 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

Supplier Data

ChIC/CUT&RUN sequencing - Anti-RUNX1 / AML1 antibody [RM1089] (AB315398)

ChIC/CUT&RUN sequencing 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 ab315398 [RM1089]. 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.