Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

Intracellular flow cytometric analysis of permeabilized Molt-4 cells using anti-RUNX1 ab92336 (red) or a rabbit IgG (negative) (green). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92336).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

Immunohistochemistry staining of RUNX1 / AML1 in formalin-fixed, paraffin-embedded Human tonsil tissue using 1/100 ab92336.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92336).

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• ICC/IF

AbReview27887****

Immunocytochemistry/ Immunofluorescence - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

ab92336 staining RUNX1 / AML1 in human glioblastoma cell line by Immunocytochemistry/ Immunofluorescence.

Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab92336 at a 1/50 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/400 dilution. Nuclei are counterstained with DAPI.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92336).

This image is courtesy of an anonymous Abreview.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

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

• IP

Lab

Immunoprecipitation - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

Lane 1 (input) : MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate, 10μg
Lane 2 (+) : MOLT-4 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab220117 in MOLT-4 whole cell lysate

ab220117 Immunoprecipitating RUNX1 / AML1 + RUNX3 + RUNX2 in MOLT-4 whole cell lysates. For western blotting, primary antibody used was ab220117 at 1 : 500 dilution (1.98 μg/ml). ab131366 VeriBlot for IP (HRP) was used for detection at 1 : 1000 dilution.

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (ab220117)

false

• IP

Lab

Immunoprecipitation - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

ab92336 (purified) at 1/50 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, ab220117 at 1/500 and HRP Veriblot for IP (ab131366) was used for detection at 1/1000 dilution.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92336).

All lanes:

Immunoprecipitation - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (<a href='/en-us/products/primary-antibodies/runx1-aml1-runx3-runx2-antibody-epr3099-ab92336'>ab92336</a>)

false

• ICC/IF

AbReview27886****

Immunocytochemistry/ Immunofluorescence - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

ab92336 staining RUNX1 / AML1 in rat glioblastoma cell line C6 by Immunocytochemistry/ Immunofluorescence.

Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab92336 at a 1/50 dilution for 16 hours at 4°C. The secondary used was a Cy3 conjugated goat anti-rabbit polyclonal used at a 1/400 dilution. Nuclei are counterstained with DAPI.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92336).

This image is courtesy of an anonymous Abreview.

• WB

Lab

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

Blocking and diluting buffer : 5% NFDM/TBST

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92336).

All lanes:

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (<a href='/en-us/products/primary-antibodies/runx1-aml1-runx3-runx2-antibody-epr3099-ab92336'>ab92336</a>) 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

false

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92336).

• WB

Lab

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] - BSA and Azide free (AB220117)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92336).

All lanes:

Western blot - Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (<a href='/en-us/products/primary-antibodies/runx1-aml1-runx3-runx2-antibody-epr3099-ab92336'>ab92336</a>) 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