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

6 product images

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  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 Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (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: Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 IP in Saos-2 whole cell lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade 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 (Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639).

  All lanes: Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639)

  Predicted band size: 57 kDa

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  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 Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (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: Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 IP in PC-3 whole cell lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade 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 (Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639).

  All lanes: Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639)

  Predicted band size: 57 kDa

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  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 Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on human osteosarcoma (PMID: 21731849) is observed. The section was incubated with Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade 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 (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 (Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639).

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  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 Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (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 Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade 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 (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 (Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639).

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  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 (Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade 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 Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 [EPR22858-106]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.
  Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

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  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 Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639 (red), or 5 μg of rabbit normal IgG Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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 (Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade ab236639).