RUNX2 Recombinant Rabbit Monoclonal Antibody [SD208-0]

RMB: 1500 特惠 1500

产品规格

50μl

100μl

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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

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

概述

产品名称

RUNX2 Recombinant Rabbit Monoclonal Antibody [SD208-0]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Recombinant protein within human 300-450.

种属反应性

Human, Mouse, Rat

验证应用

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

分子量

Predicted band size: 57 kDa

阳性对照

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.

偶联

unconjugated

克隆号

SD208-0

RRID

AB_2924311

产品特性

形态

Liquid

浓度

1ug/ul

存放说明

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

存储缓冲液

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

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

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

发表文章中的应用

WB	查看 41 篇文献如下
IHC	查看 12 篇文献如下
IF	查看 7 篇文献如下

发表文章中的种属

Mouse	查看 23 篇文献如下
Rat	查看 20 篇文献如下
Human	查看 11 篇文献如下

靶点

功能

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.

背景文献

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).

组织特异性

Specifically expressed in osteoblasts.

翻译后修饰

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.

亚细胞定位

Nucleus.

UNIPROT

SwissProt: Q13950 Human

SwissProt: Q08775 Mouse

SwissProt: Q9Z2J9 Rat

别名

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

展开

图片

☑ 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.
☑ 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.
☑ 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 (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.
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).
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) 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 (ET1612-47) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1612-47) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1612-47) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 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 1/200 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 (ET1612-47) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 ET1612-47 at 2 µg/25 µl agarose. 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 (ET1612-47) at 1/2,000 dilution. The immunostaining was performed with the IRISKit® HyperView mTSA Kit (MH900206). 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.
☑ 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.