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RUNX2 Recombinant Rabbit Monoclonal Antibody [SD208-0]

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Specification

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Catalog# ET1612-47

RUNX2 Recombinant Rabbit Monoclonal Antibody [SD208-0]

Application

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • WB

  • FC

  • IP

  • mIHC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

RUNX2 Recombinant Rabbit Monoclonal Antibody [SD208-0]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human 300-450.

Species Reactivity

Human, Mouse, Rat

Validated Applications

IF-Cell, IF-Tissue, IHC-P, WB, FC, IP, mIHC

Molecular Weight

Predicted band size: 57 kDa

Positive Control

MDA-MB-231 cell lysate, Saos-2 cell lysate, NIH/3T3 cell lysate, Saos-2, SW480, human tonsil tissue, human colon tissue, human spleen tissue, mouse bone tissue, rat maxilla tissue, C2C12.

Conjugation

unconjugated

Clone Number

SD208-0

RRID

AB_2924311

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:10,000

  • IF-Cell

  • 1:1,000-1:5,000

  • IF-Tissue

  • 1:200-1:500

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:5,000

  • IP

  • 1-2μg/sample

  • mIHC

  • 1:2,000

Target

Function

The mammalian Runt-related transcription factor (RUNX) family comprises three members, RUNX1 (also designated AML-1, PEBP2αB, CBFA2), RUNX2 (also designated AML-3, PEBP2αA, CBFA1, Osf2) and RUNX3 (also designated AML-2, PEBPαC, CBFA3). RUNX family members are DNA-binding proteins that regulate the expression of genes involved in cellular differentiation and cell cycle progression. RUNX2 is essential for skeletal mineralization in that it stimulates osteoblast differentiation of mesenchymal stem cells, promotes chondrocyte hypertrophy and contributes to endothelial cell migration and vascular invasion of developing bones. Regulating RUNX2 expression may be a useful therapeutic tool for promoting bone formation. Mutations in the C-terminus of RUNX2 are associated with cleidocranial dysplasia syndrome, an autosomal-dominant skeletal dysplasia syndrome that is characterized by widely patent calvarial sutures, clavicular hypoplasia, supernumerary teeth, and short stature.

Background References

1. Wang F et al. PTH/SDF-1a cotherapy induces CD90+CD34- stromal cells migration and promotes tissue regeneration in a rat periodontal defect model. Sci Rep 6:30403 (2016).

2. Pang J et al. ACVR1-Fc suppresses BMP signaling and chondro-osseous differentiation in an in vitro model of Fibrodysplasia ossificans progressiva. Bone 92:29-36 (2016).

Tissue Specificity

Specifically expressed in osteoblasts.

Post-translational Modification

Phosphorylated; probably by MAP kinases (MAPK). Phosphorylation by HIPK3 is required for the SPEN/MINT and FGF2 transactivation during osteoblastic differentiation (By similarity). Phosphorylation at Ser-451 by CDK1 promotes endothelial cell proliferation required for tumor angiogenesis probably by facilitating cell cycle progression. Isoform 3 is phosphorylated on Ser-340.

Subcellular Location

Nucleus.

UNIPROT

SwissProt: Q13950 Human

SwissProt: Q08775 Mouse

SwissProt: Q9Z2J9 Rat

Synonyms

Acute myeloid leukemia 3 protein antibody

Alpha subunit 1 antibody

AML3 antibody

CBF alpha 1 antibody

CBF-alpha-1 antibody

CBFA1 antibody

CCD antibody

CCD1 antibody

Cleidocranial dysplasia 1 antibody

Core binding factor antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of RUNX2 on different lysates with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/10,000 dilution.<br /><br />Lane 1: MDA-MB-231 cell lysate<br />Lane 2: Saos-2 cell lysate<br />Lane 3: LNCaP cell lysate (low expression)<br />Lane 4: NIH/3T3 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 57 kDa<br />Observed band size: 57 kDa<br /><br />Exposure time: 2 minutes 24 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of RUNX2 on different lysates with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/10,000 dilution.

    Lane 1: MDA-MB-231 cell lysate
    Lane 2: Saos-2 cell lysate
    Lane 3: LNCaP cell lysate (low expression)
    Lane 4: NIH/3T3 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 57 kDa
    Observed band size: 57 kDa

    Exposure time: 2 minutes 24 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-47) at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of Saos-2 (positive) and HeLa (negative) labeling RUNX2 with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of Saos-2 (positive) and HeLa (negative) labeling RUNX2 with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of RUNX2 on different lysates with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/5,000 dilution.<br /><br />Lane 1: Saos-2-si NT cell lysate<br />Lane 2: Saos-2-si RUNX2#1 cell lysate<br />Lane 3: Saos-2-si RUNX2#2 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 57 kDa<br />Observed band size: 57、55 kDa<br /><br />Exposure time: 43 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br /><a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a> was shown to specifically react with RUNX2 in Saos-2-si NT cells. Weakened bands were observed when Saos-2-si RUNX2 samples were tested. Saos-2-si NT and Saos-2-si RUNX2 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>, 1/5,000) and Loading control antibody (Rabbit anti-GAPDH, <a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>, 1/10,000) were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-rabbit IgG-HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of RUNX2 on different lysates with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/5,000 dilution.

    Lane 1: Saos-2-si NT cell lysate
    Lane 2: Saos-2-si RUNX2#1 cell lysate
    Lane 3: Saos-2-si RUNX2#2 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 57 kDa
    Observed band size: 57、55 kDa

    Exposure time: 43 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    ET1612-47 was shown to specifically react with RUNX2 in Saos-2-si NT cells. Weakened bands were observed when Saos-2-si RUNX2 samples were tested. Saos-2-si NT and Saos-2-si RUNX2 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1612-47, 1/5,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of C2C12 cells labeling RUNX2 with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/1,000 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of C2C12 cells labeling RUNX2 with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/1,000 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-47) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-47) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-47) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse bone tissue with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes in microwave oven. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse bone tissue with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes in microwave oven. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-47) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat maxilla tissue with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat maxilla tissue with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-47) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • RUNX2 was immunoprecipitated from 0.2 mg Saos-2 cell lysate with <a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a> at 1/10,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: Saos-2 cell lysate (input)<br />Lane 2: <a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a> IP in Saos-2 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a> in Saos-2 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 20 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    RUNX2 was immunoprecipitated from 0.2 mg Saos-2 cell lysate with ET1612-47 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1612-47 at 1/10,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: Saos-2 cell lysate (input)
    Lane 2: ET1612-47 IP in Saos-2 cell lysate
    Lane 3: Rabbit IgG instead of ET1612-47 in Saos-2 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 20 seconds; ECL: K1801

  • mIHC analysis of human tonsils tissue (Formalin/PFA-fixed paraffin-embedded sections) with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/2,000 dilution. The immunostaining was performed with the IRISKitCmTSA Kit (<a href="/products/900808" style="font-weight: bold;text-decoration: underline;">900808</a>). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

    mIHC analysis of human tonsils tissue (Formalin/PFA-fixed paraffin-embedded sections) with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/2,000 dilution. The immunostaining was performed with the IRISKitCmTSA Kit (900808). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of RUNX2 on different lysates with Rabbit anti-RUNX2 antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/5,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-RUNX2 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 57 kDa<br />Observed band size: 57 kDa<br /><br />Exposure time: 40 seconds; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of RUNX2 on different lysates with Rabbit anti-RUNX2 antibody (ET1612-47) at 1/5,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-RUNX2 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 57 kDa
    Observed band size: 57 kDa

    Exposure time: 40 seconds; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-47) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Flow cytometric analysis of Saos-2 cells labeling RUNX2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1612-47" style="font-weight: bold;text-decoration: underline;">ET1612-47</a>, red) at 1/5,000 dilution and competitor's antibody (red) at 1/2,000 dilution, compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Saos-2 cells labeling RUNX2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-47, red) at 1/5,000 dilution and competitor's antibody (red) at 1/2,000 dilution, compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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