Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free (AB264077)

Immunohistochemical analysis of paraffin-embedded human osteosarcoma tissue labeling RUNX2 with ab236639 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human osteosarcoma (PMID : 21731849) is observed. The section was incubated with ab236639 for 15 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab236639).

• IP

Unknown

Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free (AB264077)

RUNX2 was immunoprecipitated from 0.35 mg PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate 10ug with ab236639 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab236639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2 : ab236639 IP in PC-3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab236639 in PC-3 whole cell lysate

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

Exposure time : 30 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab236639).

All lanes:

Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (<a href='/en-us/products/primary-antibodies/runx2-antibody-epr22858-106-chip-grade-ab236639'>ab236639</a>)

Predicted band size: 57 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free (AB264077)

RUNX2 was immunoprecipitated from 0.35 mg Saos-2 (human osteosarcoma epithelial) whole cell lysate with ab236639 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab236639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Saos-2 (human osteosarcoma epithelial) whole cell lysate 10ug

Lane 2 : ab236639 IP in Saos-2 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab236639 in Saos-2 whole cell lysate

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

Exposure time : 30 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab236639).

All lanes:

Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (<a href='/en-us/products/primary-antibodies/runx2-antibody-epr22858-106-chip-grade-ab236639'>ab236639</a>)

Predicted band size: 57 kDa

false

• ChIP

Unknown

ChIP - Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free (AB264077)

Chromatin was prepared from Saos-2 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.

The ChIP was performed with 25 μg of chromatin, 5 μg of ab236639 (red), or 5 μg of rabbit normal IgG ab172730 (gray) and 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab236639).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free (AB264077)

Immunohistochemical analysis of paraffin-embedded rat embryo 14.5 day tissue labeling RUNX2 with ab236639 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the cartilage cells in the rat embryo 14.5 day tissue is observed. The section was incubated with ab236639 for 15 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab236639).

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free (AB264077)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab236639). ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5X10^5 of positive cell line Saos-2 or low expression cell line HeLa were used along with 5µg of ab236639 [EPR22858-106]. 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.