rabbit antibodies against runx3  (Proteintech)

Journal: Journal of Oncology

Article Title: Oncosuppressive Role of RUNX3 in Human Astrocytomas

doi: 10.1155/2019/1232434

Figure Lengend Snippet: (a) Representative results for methylation-specific PCR of RUNX3 gene in different malignancy grade gliomas. M: PCR with primers for methylated RUNX3. U: PCR with primers for unmethylated RUNX3. Positive control: bisulfite converted universal methylated human DNA standard (Zymo Research, USA). Negative control: bisulfite converted normal lymphocyte DNA. Normal: “normal human brain DNA” (Zymo Research, cat. no. D5018). Arrows indicate methylated allele. (b) RUNX3 promoter methylation frequency in different astrocytoma grade. GBM revealed significant increase in RUNX3 gene promoter methylation frequency compared to I-III WHO grade astrocytomas (p<0.001, chi-square test). (c) Kaplan-Meier curves for survival (months) of all glioma patients (n=136) stratified by RUNX3 gene methylation status revealed significantly shorter survival of patients with RUNX3 methylated tumours (Log-rank test, χ 2 =44.68, df=1, p<0.001).

Article Snippet: Primary rabbit antibody against RUNX3 (Antibodies-Online, cat no. ABIN739370) diluted 1:500 in 5% nonfat milk in PBS was used for RUNX3 protein detection applying 2-hour incubation on a platform shaker at room temperature.

Techniques: Methylation, Positive Control, Negative Control

Journal: Journal of Oncology

Article Title: Oncosuppressive Role of RUNX3 in Human Astrocytomas

doi: 10.1155/2019/1232434

Figure Lengend Snippet: Associations between grade I-IV astrocytoma patients' clinical data and RUNX3 gene molecular properties.

Article Snippet: Primary rabbit antibody against RUNX3 (Antibodies-Online, cat no. ABIN739370) diluted 1:500 in 5% nonfat milk in PBS was used for RUNX3 protein detection applying 2-hour incubation on a platform shaker at room temperature.

Techniques: Methylation, Expressing

Journal: Journal of Oncology

Article Title: Oncosuppressive Role of RUNX3 in Human Astrocytomas

doi: 10.1155/2019/1232434

Figure Lengend Snippet: (a) Representative Western blot result of RUNX3 protein expression in astrocytomas. AI-AIII: astrocytoma malignancy grades I-III, respectively; GBM: glioblastoma. Two isoforms of RUNX3 were identified in all the specimens that is consistent with what has been described in the literature . (b) RUNX3 protein expression levels in different astrocytoma grade. Protein expression was significantly downregulated in glioblastomas (GBM) as compared to grade II astrocytomas (p<0.005, Kruskal-Wallis test) and a tendency as compared to grade III astrocytomas (p=0.124, Kruskal-Wallis test). (c) Relative RUNX3 protein expression stratified by promoter methylation groups. Significant association between RUNX3 gene methylation and protein expression was found (p=0.026, Kruskal-Wallis test). (d) Kaplan-Meier curves for survival of all astrocytoma patients (n=72) stratified in two groups (low; high) according to protein expression revealed significant better survival rates for patient with high RUNX3 protein level (Log-rank test, χ 2=6.11, df=1, p=0.013). (e) Kaplan-Meier survival curves of high malignancy grade (III-IV) astrocytoma patients only (n=39) (Log-rank test, χ 2=13.74, df=1, p<0.001).

Article Snippet: Primary rabbit antibody against RUNX3 (Antibodies-Online, cat no. ABIN739370) diluted 1:500 in 5% nonfat milk in PBS was used for RUNX3 protein detection applying 2-hour incubation on a platform shaker at room temperature.

Techniques: Western Blot, Expressing, Methylation

Journal: Journal of Oncology

Article Title: Oncosuppressive Role of RUNX3 in Human Astrocytomas

doi: 10.1155/2019/1232434

Figure Lengend Snippet: Multivariate Cox regression analysis applying Backward Conditional method.

Article Snippet: Primary rabbit antibody against RUNX3 (Antibodies-Online, cat no. ABIN739370) diluted 1:500 in 5% nonfat milk in PBS was used for RUNX3 protein detection applying 2-hour incubation on a platform shaker at room temperature.

Techniques: Methylation

Journal: Journal of Oncology

Article Title: Oncosuppressive Role of RUNX3 in Human Astrocytomas

doi: 10.1155/2019/1232434

Figure Lengend Snippet: (a) WB of RUNX3 protein expression after U87-MG cells transfection with pcDNA3-RUNX3 construct. NTC: nontransfected cells lysate; 1: cells transfected with 100 ng of pcDNA3-RUNX3 vector; 2: cells transfected with 50 ng of pcDNA3-RUNX3 vector. Equal amount of total protein extract (60 μ g) was loaded per each gel lane. The same amount of cell (b) RUNX3 promoter methylation status in U87-MG cells. (c) Results from glioblastoma U87-MG cells viability assay applying MTT test after 24 h of 100 ng of DNA transfection. RUNX3: RUNX3 gene in pcDNA3 expression vector (pcDNA3-RUNX3); GFP: green fluorescing protein in pcDNA4TO expression vector (pcDNA4TO-GFP); pcDNA3: empty (control) vector; NTC: nontransfected cell control. Overexpression of RUNX3 significantly decreased U87-MG cell viability starting from 50 ng of vector used as compared to the cells transfected with GFP vector. Moreover even bigger effect of decreased cell viability was obtained when 100 ng of RUNX3 was transfected.

Article Snippet: Primary rabbit antibody against RUNX3 (Antibodies-Online, cat no. ABIN739370) diluted 1:500 in 5% nonfat milk in PBS was used for RUNX3 protein detection applying 2-hour incubation on a platform shaker at room temperature.

Techniques: Expressing, Transfection, Construct, Plasmid Preparation, Methylation, Viability Assay, Over Expression

Journal: Development (Cambridge, England)

Article Title: Termination of cell-type specification gene programs by the miR-183 cluster determines the population sizes of low-threshold mechanosensitive neurons.

doi: 10.1242/dev.165613

Figure Lengend Snippet: Fig. 1. The population sizes of TRKB+ and TRKC+ neurons are altered in the P0 miRCKO mice. (A-C) The percentage of TRKB+ neurons was increased in the miRCKO mice. Immunostaining for TRKB on DRG sections of control Wnt1-Cre mice (A) and the miRCKO mice (B) as well as quantification of the percentage of TRKB+ neurons in L3-L5 ganglions (C, n=3, 4). (D) The number of total neurons (ISL1+) in L5 DRG was not altered in the miRCKO mice compared with controls (n=3, 4; not significant). (E-G) The percent of total TRKC neurons were decreased in the miRCKO mice. Double immunostaining for TRKC and RUNX3 on DRG sections of Wnt1-Cre mice (E) and miR CKO mice (F), arrow heads in E and F point to NF3 (TRKC+RUNX3-) neurons, inset are higher magnifications of areas outlined in E and F. (G) Quantification of the percentage of total TRKC, TRKC+RUNX3+ (not significant) and TRKC+RUNX3- neurons in L3-L5 ganglions (n=4, 4). Data shown in mean±s.e.m. and analyzed by t-test and P<0.05 is statistically significant. *P<0.05, **P<0.01. Scale bar =µ.

Article Snippet: The following primary antibodies were used: mouse antibodies against ISL1 (DSHB, USA, 1:100), SHOX2 (Santa Cruz, USA, 1:400), NFH (neurofilament, heavy polypeptide) (CloneN52; Sigma-aldrich, St. Louis, MO, USA, 1:500); rabbit antibodies against RUNX3 (gifts from Thomas Jessell, Columbia University Medical Center, 1:300), ISL1 (gifts from Thomas Jessell, Columbia University Medical Center, 1:250), NFH (Millipore, USA, 1:200), TRKA (Millipore, USA, 1:500), TRKC (Cell signaling, USA, 1:500), NECAB2 (Proteintech Europe, 1:1000), CALB1 (Millipore, USA, 1:500) and chicken-TRKB (kind gift from Louis F. Reichardt, 1:2000); goat antibodies against TRKA (R&D systems, Minneapolis, MN, USA, 1:500), TRKB (R&D systems, Minneapolis, MN, USA, 1:500), TRKC (R&D, 1:500), Ret (R&D, 1:100) and GFP (Abcam, USA, 1:500); Guinea pig antibodies against TLX3 (kind gift from Carmen Birchmeier).

Techniques: Immunostaining, Control, Double Immunostaining

Journal: Molecular Medicine Reports

Article Title: Loss of RUNX3 is significantly associated with advanced tumor grade and stage in endometrial cancers

doi: 10.3892/mmr.2018.8915

Figure Lengend Snippet: Expression of RUNX3 according to clinicopathological factors.

Article Snippet: To block endogenous peroxidase, the sections were incubated in 3% hydrogen peroxide in methanol at room temperature for 20 min. Next, sections were washed three times with distilled water, soaked in PBS pH 7.4 for 3 min, and then incubated with rabbit polyclonal antibody against human RUNX3 (39301; 1:100; Active Motif, Inc., Carlsbad, CA, USA) at room temperature for 1 h. Sections were washed with PBS three times for 3 min each and then were incubated with secondary antibody EnVision HRP-Labelled Polymer anti-rabbit (K4002; Dako; Agilent Technologies, Inc., Santa Clara, CA, USA) for 30 min at room temperature.

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: Loss of RUNX3 is significantly associated with advanced tumor grade and stage in endometrial cancers

doi: 10.3892/mmr.2018.8915

Figure Lengend Snippet: (A) Expression patterns of RUNX3 mRNA in two normal endometrial tissues, two endometrial cancer tissues, and two endometrial cancer cell lines, as determined by reverse-transcription-PCR. GAPDH was used as a standard reference (B) Methylation-specific PCR for the two endometrial cancer cell lines, HEC1-α and Ishikawa. RUNX3, runt-related transcription factor 3; PCR, polymerase chain reaction; N, normal endometrial tissue; T, endometrial cancer tissue; M, methylated; U, unmethylated; PC, positive control; NC, negative control.

Article Snippet: To block endogenous peroxidase, the sections were incubated in 3% hydrogen peroxide in methanol at room temperature for 20 min. Next, sections were washed three times with distilled water, soaked in PBS pH 7.4 for 3 min, and then incubated with rabbit polyclonal antibody against human RUNX3 (39301; 1:100; Active Motif, Inc., Carlsbad, CA, USA) at room temperature for 1 h. Sections were washed with PBS three times for 3 min each and then were incubated with secondary antibody EnVision HRP-Labelled Polymer anti-rabbit (K4002; Dako; Agilent Technologies, Inc., Santa Clara, CA, USA) for 30 min at room temperature.

Techniques: Expressing, Reverse Transcription, Methylation, Polymerase Chain Reaction, Positive Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: Loss of RUNX3 is significantly associated with advanced tumor grade and stage in endometrial cancers

doi: 10.3892/mmr.2018.8915

Figure Lengend Snippet: Immunohistochemical staining of endometrial cancer tissues for RUNX3. (A) Grade 3 endometrial cancer sample displays negative staining (0). (B) Grade 2 endometrial cancer sample displays positive +1 staining (C) Grade 1 endometrial cancer displays +2 positive staining. (D) Grade 1 endometrial cancer displays +2 positive staining. Representative images at magnification, ×200. RUNX3, runt-related transcription factor 3.

Article Snippet: To block endogenous peroxidase, the sections were incubated in 3% hydrogen peroxide in methanol at room temperature for 20 min. Next, sections were washed three times with distilled water, soaked in PBS pH 7.4 for 3 min, and then incubated with rabbit polyclonal antibody against human RUNX3 (39301; 1:100; Active Motif, Inc., Carlsbad, CA, USA) at room temperature for 1 h. Sections were washed with PBS three times for 3 min each and then were incubated with secondary antibody EnVision HRP-Labelled Polymer anti-rabbit (K4002; Dako; Agilent Technologies, Inc., Santa Clara, CA, USA) for 30 min at room temperature.

Techniques: Immunohistochemical staining, Staining, Negative Staining

Journal: Molecular Medicine Reports

Article Title: Loss of RUNX3 is significantly associated with advanced tumor grade and stage in endometrial cancers

doi: 10.3892/mmr.2018.8915

Figure Lengend Snippet: Methylation-specific polymerase chain reaction analysis of the RUNX3 promoter was performed in six endometrial cancer tissues (T1-T6) and eight normal endometrial tissues (N1-N8). Positive findings were observed in T1-T4 and N4 samples. RUNX3, runt-related transcription factor 3; SM, size marker; C, control; M, methylated; U, unmethylated.

Article Snippet: To block endogenous peroxidase, the sections were incubated in 3% hydrogen peroxide in methanol at room temperature for 20 min. Next, sections were washed three times with distilled water, soaked in PBS pH 7.4 for 3 min, and then incubated with rabbit polyclonal antibody against human RUNX3 (39301; 1:100; Active Motif, Inc., Carlsbad, CA, USA) at room temperature for 1 h. Sections were washed with PBS three times for 3 min each and then were incubated with secondary antibody EnVision HRP-Labelled Polymer anti-rabbit (K4002; Dako; Agilent Technologies, Inc., Santa Clara, CA, USA) for 30 min at room temperature.

Techniques: Methylation, Polymerase Chain Reaction, Marker, Control

Journal: Molecular Medicine Reports

Article Title: Loss of RUNX3 is significantly associated with advanced tumor grade and stage in endometrial cancers

doi: 10.3892/mmr.2018.8915

Figure Lengend Snippet: Expression and methylation of RUNX3 according to clinicopathological factors.

Article Snippet: To block endogenous peroxidase, the sections were incubated in 3% hydrogen peroxide in methanol at room temperature for 20 min. Next, sections were washed three times with distilled water, soaked in PBS pH 7.4 for 3 min, and then incubated with rabbit polyclonal antibody against human RUNX3 (39301; 1:100; Active Motif, Inc., Carlsbad, CA, USA) at room temperature for 1 h. Sections were washed with PBS three times for 3 min each and then were incubated with secondary antibody EnVision HRP-Labelled Polymer anti-rabbit (K4002; Dako; Agilent Technologies, Inc., Santa Clara, CA, USA) for 30 min at room temperature.

Techniques: Expressing, Methylation

Journal: Molecular Medicine Reports

Article Title: Loss of RUNX3 is significantly associated with advanced tumor grade and stage in endometrial cancers

doi: 10.3892/mmr.2018.8915

Figure Lengend Snippet: Restoration of RUNX3 mRNA expression in HEC1-α cells following ADC treatment. Lane 1, 0 µM ADC control; lane 2, 0.5 µM ADC; lane 3, 1 µM ADC; lane 4, 5 µM ADC; lane 5, no template control. RUNX3, runt-related transcription factor 3; ADC, 5-aza-2′-deoxycytidine.

Article Snippet: To block endogenous peroxidase, the sections were incubated in 3% hydrogen peroxide in methanol at room temperature for 20 min. Next, sections were washed three times with distilled water, soaked in PBS pH 7.4 for 3 min, and then incubated with rabbit polyclonal antibody against human RUNX3 (39301; 1:100; Active Motif, Inc., Carlsbad, CA, USA) at room temperature for 1 h. Sections were washed with PBS three times for 3 min each and then were incubated with secondary antibody EnVision HRP-Labelled Polymer anti-rabbit (K4002; Dako; Agilent Technologies, Inc., Santa Clara, CA, USA) for 30 min at room temperature.

Techniques: Expressing, Control

Journal: PLoS ONE

Article Title: RUNX3 Mediates Suppression of Tumor Growth and Metastasis of Human CCRCC by Regulating Cyclin Related Proteins and TIMP-1

doi: 10.1371/journal.pone.0032961

Figure Lengend Snippet: a and b. Representative immunohistochemical photographs were taken at different magnifications in CCRCC tumor tissues and their matched noncancerous counterparts (×100 a1, ×200 a2, ×400 a3 and negative control a4 for noncancerous tissues; ×100 b1, ×200 b2, ×400 b3 and negative control b4 for tumor tissues). c. Expression protein levels of Runx3 in six CCRCC and the matched adjacent noncancerous tissues. d.Expression mRNA levels of RUNX3 in the CCRCC-derived cell lines and human kidney proximal tubular cell lines by real time RT-PCR. 18S was used as an internal control. * P <0.05 vs HKC cells. e. Expression protein levels of RUNX3 in the CCRCC-derived cell lines and human kidney proximal tubular cell lines by Western Blot. Tubulin was used as an internal control.

Article Snippet: Immunohistochemistry was done as described using a rabbit polyclonal antibody against human RUNX3 at a dilution of 1∶200 (Active Motif, Carlsbad, CA) .

Techniques: Immunohistochemical staining, Negative Control, Expressing, Derivative Assay, Quantitative RT-PCR, Control, Western Blot

Journal: PLoS ONE

Article Title: RUNX3 Mediates Suppression of Tumor Growth and Metastasis of Human CCRCC by Regulating Cyclin Related Proteins and TIMP-1

doi: 10.1371/journal.pone.0032961

Figure Lengend Snippet: Clinicopathological associations of RUNX3 expression in patients with CCRCC.

Article Snippet: Immunohistochemistry was done as described using a rabbit polyclonal antibody against human RUNX3 at a dilution of 1∶200 (Active Motif, Carlsbad, CA) .

Techniques: Expressing

Journal: PLoS ONE

Article Title: RUNX3 Mediates Suppression of Tumor Growth and Metastasis of Human CCRCC by Regulating Cyclin Related Proteins and TIMP-1

doi: 10.1371/journal.pone.0032961

Figure Lengend Snippet: a. After infection, the expression of RUNX3 in 786-O-Ctrl and 786-O-RUNX3 was evaluated by Western blot. Tubulin was used as an internal control. b. Effect of RUNX3 in regulating 786-O cells proliferation. Monolayer growth rates of 786-O, 786-O-Ctrl and 786-O-RUNX3 cells were determined by MTT assays. Values represent the mean (SEM) from at least three separate experiments. * P <0.05 vs 786-O cells. c. Effect of RUNX3 on colony formation of 786-O cells. Cells were placed in media containing soft agar and incubated for 17 days. The number of foci >100 µm was counted. Values represent the mean (SEM) from at least three separate experiments, each conducted in triplicate. * P <0.05 vs 786-O-Ctrl cells. d. Cell migration assays. Representative fields of migration cells on the membrane (magnification of ×200). Average migration cell number per field. The migration cell number of 786-O-RUNX3 is drastically decreased than that transfected with negative control. * P <0.05 vs 786-O-Ctrl cells, Student’s T-test, n = 10.

Article Snippet: Immunohistochemistry was done as described using a rabbit polyclonal antibody against human RUNX3 at a dilution of 1∶200 (Active Motif, Carlsbad, CA) .

Techniques: Infection, Expressing, Western Blot, Control, Incubation, Migration, Membrane, Transfection, Negative Control

Journal: PLoS ONE

Article Title: RUNX3 Mediates Suppression of Tumor Growth and Metastasis of Human CCRCC by Regulating Cyclin Related Proteins and TIMP-1

doi: 10.1371/journal.pone.0032961

Figure Lengend Snippet: a. Average tumor weight was measurement of the excised tumors at the time of sacrifice. * P <0.05 vs 786-O-Ctrl cells. b. Average tumor size was estimated by physical measurement of the excised tumor at different time. * P <0.05 vs 786-O-Ctrl cells. d. 786-O-Ctrl cells and 786-O-RUNX3 cells were cultured in DMEM for 24 h. Cells were harvested and processed for FACS analysis.

Article Snippet: Immunohistochemistry was done as described using a rabbit polyclonal antibody against human RUNX3 at a dilution of 1∶200 (Active Motif, Carlsbad, CA) .

Techniques: Cell Culture

Journal: PLoS ONE

Article Title: RUNX3 Mediates Suppression of Tumor Growth and Metastasis of Human CCRCC by Regulating Cyclin Related Proteins and TIMP-1

doi: 10.1371/journal.pone.0032961

Figure Lengend Snippet: a. The expression of cyclin D1, cyclin E, cdk2, cdk4, p-Rb, Rb, p27, MMP2, MMP9, TIMP-1 and TIMP-2 proteins were evaluated in 786-O-Ctrl and 786-O-RUNX3 cells by Western blot. b. The expression of cyclin D1, cyclin E, cdk2, cdk4, p-Rb, Rb, p27, MMP2, MMP9, TIMP-1 and TIMP-2 proteins were evaluated in HKC-Ctrl and HKC-siRUNX3 by Western blot. All examined gene expression levels quantitatively analyzed and expressed as the ratios over β-actin.

Article Snippet: Immunohistochemistry was done as described using a rabbit polyclonal antibody against human RUNX3 at a dilution of 1∶200 (Active Motif, Carlsbad, CA) .

Techniques: Expressing, Western Blot, Gene Expression