RUNX2 Recombinant Rabbit Monoclonal Antibody [SD208-0] - BSA and Azide free
Catalog# HA750295
RUNX2 Recombinant Rabbit Monoclonal Antibody [SD208-0] - BSA and Azide free
-
IF-Cell
-
IF-Tissue
-
IHC-P
-
WB
-
FC
-
IP
-
Human
-
Mouse
-
Rat
-
ET1612-47
含抗保成分
-
unconjugated
概述
产品名称
RUNX2 Recombinant Rabbit Monoclonal Antibody [SD208-0] - BSA and Azide free
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
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
产品特性
形态
Liquid
浓度
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*PBS (pH7.4).
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
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.
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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 (HA750295) 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 HA750295 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750295 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: HA750295 IP in Saos-2 cell lysate
Lane 3: Rabbit IgG instead of HA750295 in Saos-2 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 20 seconds; ECL: K1801 -
☑ Knockdown (KD)
Western blot analysis of RUNX2 on different lysates with Rabbit anti-RUNX2 antibody (HA750295) 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 (HA750295) 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.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750295, 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).
请注意: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
浙公网安备 33019202000643号