Anti-RUNX2 抗体 [EPR14334]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR14334] (AB192256)

Immunohistochemical analysis of paraffin-embedded Human osteosarcoma tissue labeling RUNX2 with ab192256 at 1/1000 dilution. A ready to use HRP Polymer for Rabbit IgG was used as the secondary. Hematoxylin counterstain.

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-RUNX2 antibody [EPR14334] (AB192256)

Immunofluorescent analysis of 4% formaldehyde fixed PC3 cells labeling RUNX2 using ab192256 at a 1/500 dilution. A Goat anti rabbit IgG (Alexa Fluor®488) ab150077 was used as the secondary at a 1/200 dilution. Counterstain DAPI. Permeabilized using 0.1% Triton X-100. The two negative controls : 1. Primary ab concentration (anti-RUNX2) is 1 : 500 dilution, Secondary ab (Goat anti mouse IgG (Alexa Fluor®594)) is 1 : 500 dilution; 2. Primary ab concentration (anti-RUNX2) is 1 : 500 dilution, Secondary ab (Goat anti mouse IgG (Alexa Fluor®594)) is 1 : 500 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR14334] (AB192256)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling RUNX2 with ab192256 at 1/1000 dilution. A ready to use HRP Polymer for Rabbit IgG was used as the secondary. Hematoxylin counterstain.

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

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RUNX2 antibody [EPR14334] (AB192256)

ab192256 staining RUNX2 in PC-3 (human prostate adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/50. A goat anti rabbit IgG (Alexa Fluorr^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RUNX2 antibody [EPR14334] (AB192256)

Immunocytochemistry/Immunofluorescence analysis of Saos-2 (Human osteosarcoma cell line) labeling RUNX2 with purified ab192256 at 1/1000 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor^®488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR14334] (AB192256)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling RUNX2 with ab192256 at 1/1000 dilution. A ready to use HRP Polymer for Rabbit IgG was used as the secondary. Hematoxylin counterstain.

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

• WB

Lab

Western blot - Anti-RUNX2 antibody [EPR14334] (AB192256)

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

ab181602 was used as a GAPDH loading control

Below the target band, there are multi bands may be caused by protein degradation. We suggest customer to use freshly lysate to minimize protein degradation in western blot.
Low express : MCF-7 (PMID : 32509221, 16166639, 20591170)

All lanes:

Western blot - Anti-RUNX2 antibody [EPR14334] (ab192256) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 6:

C2C12 (Mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 7:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60 kDa

false

Exposure time: 10s

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-RUNX2 antibody [EPR14334] (AB192256)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10⁵ Saos-2 cells and 5 µg of ab192256 [EPR14334]. 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-RUNX2 antibody [EPR14334] (AB192256)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10⁵ Saos-2 cells and 5 µg of ab192256 [EPR14334]. 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-RUNX2 antibody [EPR14334] (AB192256)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10⁵ Saos-2 cells and 5 µg of ab192256 [EPR14334]. 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.

• WB

Lab

Western blot - Anti-RUNX2 antibody [EPR14334] (AB192256)

Western blot : Anti-RUNX2 antibody [EPR14334] (ab192256) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab192256 was shown to bind specifically to RUNX2. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

All lanes:

Western blot - Anti-RUNX2 antibody [EPR14334] (ab192256)

Lane 1:

Saos-2 cell lysate

Lane 2:

MC3T3-E1 undifferentiated cell lysate

Lane 3:

MC3T3-E1 7-day Osteogenic differentiation cell lysate

Lane 4:

MC3T3-E1 14-day Osteogenic differentiation cell lysate

Lane 5:

MC3T3-E1 28-day Osteogenic differentiation cell lysate

Lane 6:

C2C12 cell lysate

Lane 7:

SH-SY5Y cell lysate

Lane 8:

NIH/3T3 cell lysate

Lane 9:

LNCaP cell lysate

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