Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (AB92336)

Immunohistochemistry staining of RUNX1 / AML1 in formalin-fixed, paraffin-embedded Human tonsil tissue using 1/100 ab92336. Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (AB92336)

IHC image of RUNX1 / AML1 + RUNX3 + RUNX2 staining in a section of frozen normal human tonsil performed on a Leica BOND^TM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab92336, 1/500 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (AB92336)

Intracellular flow cytometric analysis of permeabilized Molt-4 cells using anti-RUNX1 ab92336 (red) or a rabbit IgG (negative) (green).

• IP

Lab

Immunoprecipitation - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (AB92336)

ab92336 (purified) at 1/20 immunoprecipitating RUNX1 / AML1 + RUNX3 + RUNX2 in 10 μg Molt-4 (Human lymphoblastic leukemia T lymphoblast）whole cell lysate (Lanes 1 and 2, observed at 49 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730) instead of ab92336 in Molt-4 whole cell lysate. For western blotting, ab92336 at 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336)

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• WB

Lab

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (AB92336)

Blocking and diluting buffer : 5% NFDM/TBST

Exposure time : 10 seconds and 100 seconds

All lanes:

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1/1000 dilution

Lane 1:

Myc-DDK-tagged Human RUNX1 recombinant protein at 20 ng

Lane 2:

His-tagged Human RUNX2 recombinant protein at 20 ng

Lane 3:

His-tagged Human RUNX3 recombinant protein at 20 ng

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

Predicted band size: 48.7 kDa

Observed band size: 48 kDa,55 kDa

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• WB

Unknown

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (AB92336)

All lanes:

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1/10000 dilution

Lane 1:

MOLT4 cell lysate at 10 µg

Lane 2:

fetal thymus lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

false

• WB

Lab

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (AB92336)

Blocking/Diluting buffer and concentration : 5% NFDM /TBST

All lanes:

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1.28 µg/mL

Lane 1:

Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

Molt-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 3:

WEHI-3 (Mouse leukemia lymphoblast) whole cell lysate at 20 µg

Lane 4:

Mouse thymus lysate at 20 µg

Lane 5:

CTLL-2 (Mouse T lymphocyte) whole cell lysate at 20 µg

Lane 6:

Rat thymus 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 0.05 µg/mL

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• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (AB92336)

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 5µg of ab92336 [EPR3099]. 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. Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• WB

Lab

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (AB92336)

Blocking/Diluting buffer and concentration : 5% NFDM /TBST

All lanes:

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1.28 µg/mL

Lane 1:

Mouse spleen lysate at 20 µg

Lane 2:

Mouse thymus 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 0.05 µg/mL

false