arhgap29 primary antibody  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: Arhgap29 Deficiency Directly Leads to Systemic and Craniofacial Skeletal Abnormalities

doi: 10.3390/ijms26104647

Figure Lengend Snippet: General characteristics of Arhgap29 −/− mice. ( A ) Gross image of the E15.5 mice. ( B ) Image of the head of the P0 mice (the black dashed box indicates the mandible). The penetrance of mandibular anomalies in Arhgap29 −/− mice is 30% (3/10). ( C ) Image of the palate of the E17.5 mice. The black dashed line indicates the cleft palate with 31.25% penetrance (5/16). ( D ) Image of the forelimb digits of the E14.5 mice. The black arrow indicates ectrodactyly with 96.67% penetrance (29/30).

Article Snippet: After dewaxing and rehydration, the sections were incubated with the Arhgap29 primary antibody (sc-377022, Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C, followed by incubation with a secondary antibody (Bioss, Beijing, China) for 30 min at 37 °C.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Arhgap29 Deficiency Directly Leads to Systemic and Craniofacial Skeletal Abnormalities

doi: 10.3390/ijms26104647

Figure Lengend Snippet: ( A ) Three-dimensional reconstruction images of micro-CT scanning of WT and Arhgap29 deletion specimens. The white dashed line indicates the outline of the collapsed skull of the Arhgap29 −/− mice. FR, frontal bone; PA, parietal bone; IP, interparietal bone; M, mandible; co, coronal suture; sa, sagittal suture; la, lambdoidal suture. ( B ) The quantitative analysis of the cranium and mandible showed that the average bone surface area, bone volume, and bone mineral content were reduced to varying degrees in both the cranium and mandible of Arhgap29 −/− mice compared with WT mice. BV, bone volume; BS, bone surface; BMC, bone mineral content. All data: *** p < 0.001; ** p < 0.01; * p < 0.05.

Article Snippet: After dewaxing and rehydration, the sections were incubated with the Arhgap29 primary antibody (sc-377022, Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C, followed by incubation with a secondary antibody (Bioss, Beijing, China) for 30 min at 37 °C.

Techniques: Micro-CT

Journal: International Journal of Molecular Sciences

Article Title: Arhgap29 Deficiency Directly Leads to Systemic and Craniofacial Skeletal Abnormalities

doi: 10.3390/ijms26104647

Figure Lengend Snippet: Alcian blue and Alcian blue/alizarin red staining results of cartilage and bone in the craniofacial region and limbs of WT and Arhgap29 −/− mice. ( A ) Alcian blue staining was performed on Meckel’s cartilage during three developmental stages: E13.5, E14.5, and P0. ( B ) Alcian blue/alizarin red staining of cranial and facial bones at P0. ( C ) Alcian blue/alizarin red staining of the mandible at P0. ( D ) Alcian blue/alizarin red staining of the whole skeleton of mice. ( E ) Alcian blue/alizarin red staining of the front limbs of mice; the Arhgap29 −/− mice have ectrodactyly.

Article Snippet: After dewaxing and rehydration, the sections were incubated with the Arhgap29 primary antibody (sc-377022, Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C, followed by incubation with a secondary antibody (Bioss, Beijing, China) for 30 min at 37 °C.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Arhgap29 Deficiency Directly Leads to Systemic and Craniofacial Skeletal Abnormalities

doi: 10.3390/ijms26104647

Figure Lengend Snippet: Histological characteristics of Meckel’s cartilage. The results of H&E staining of Meckel’s cartilage in E13.5 ( A ), E15.5 ( B ), and E17.5 ( C ) mice, along with Alcian blue staining of Meckel’s cartilage in P0 mice ( D ). ( A – C ) illustrate the coronal section staining results of the mouse head, demonstrating the delayed hypertrophy of Meckel’s chondrocytes in Arhgap29 −/− mice. ( D ) presents the axial section staining results of the mouse head, which reveal the delayed degeneration of Meckel’s cartilage. The red dashed line indicates Meckel’s cartilage (MC); the mandible is labeled as M. Three mice from each group were chosen for every time period.

Article Snippet: After dewaxing and rehydration, the sections were incubated with the Arhgap29 primary antibody (sc-377022, Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C, followed by incubation with a secondary antibody (Bioss, Beijing, China) for 30 min at 37 °C.

Techniques: Staining, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Arhgap29 Deficiency Directly Leads to Systemic and Craniofacial Skeletal Abnormalities

doi: 10.3390/ijms26104647

Figure Lengend Snippet: Experimental results of osteogenesis in mandibular tissue. ( A , B ) Von Kossa staining of mandibular tissue sections from E17.5 mice. ( C ) ALP staining of mandibular tissue sections from E17.5 WT mice. ALP-positive cells (shown in blue) are distributed on the surface of the bone matrix. ( D ) Immunohistochemical staining results of mandibular tissue sections from E17.5 WT mice. Arhgap29 -positive cells (shown in brown-yellow) are located on the surface of the bone matrix. (The red arrow indicates osteoblasts).

Article Snippet: After dewaxing and rehydration, the sections were incubated with the Arhgap29 primary antibody (sc-377022, Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C, followed by incubation with a secondary antibody (Bioss, Beijing, China) for 30 min at 37 °C.

Techniques: Staining, Immunohistochemical staining

Journal: International Journal of Molecular Sciences

Article Title: Arhgap29 Deficiency Directly Leads to Systemic and Craniofacial Skeletal Abnormalities

doi: 10.3390/ijms26104647

Figure Lengend Snippet: Experimental results of osteoclast activity in mouse mandibular tissue. ( A , B ) TRAP staining of mandibles in E17.5 WT and Arhgap29 −/− mice (osteoclasts are stained red, as indicated by the black arrows). ( C ) Staining of osteoclast marker TRAP in mandibles of WT mice at E17.5. ( D ) Immunohistochemical staining of osteoclasts in mandibles of WT mice at E17.5 (with positive cells indicated in brownish-yellow).

Article Snippet: After dewaxing and rehydration, the sections were incubated with the Arhgap29 primary antibody (sc-377022, Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C, followed by incubation with a secondary antibody (Bioss, Beijing, China) for 30 min at 37 °C.

Techniques: Activity Assay, Staining, Marker, Immunohistochemical staining

Journal: International Journal of Molecular Sciences

Article Title: Arhgap29 Deficiency Directly Leads to Systemic and Craniofacial Skeletal Abnormalities

doi: 10.3390/ijms26104647

Figure Lengend Snippet: Analysis of transcriptome sequencing results for E17.5 mandibular tissue. ( A ) Volcano plot of differentially expressed genes. ( B ) GO classification annotation and enrichment analysis. ( C ) KEGG classification annotation and pathway enrichment analysis. ( D ) Heatmap of differentially expressed gene clustering in calcium signaling pathway. ( E ) Heatmap of differentially expressed gene clustering in cell differentiation. ( F ) qPCR validation of calcium signaling pathway-related molecules. ( G ) qPCR validation of cell differentiation-related molecules. (The internal reference gene used was Gapdh , and the relative expression of the Arhgap29 −/− group was calculated based on the gene expression levels of the WT group. Statistical analyses of differences were performed using a t -test. * indicates a statistically significant difference between groups. *** p < 0.001; ** p < 0.01; ns indicates no significant difference.)

Article Snippet: After dewaxing and rehydration, the sections were incubated with the Arhgap29 primary antibody (sc-377022, Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C, followed by incubation with a secondary antibody (Bioss, Beijing, China) for 30 min at 37 °C.

Techniques: Sequencing, Cell Differentiation, Biomarker Discovery, Expressing, Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: Arhgap29 Deficiency Directly Leads to Systemic and Craniofacial Skeletal Abnormalities

doi: 10.3390/ijms26104647

Figure Lengend Snippet: In vitro cell experiment results. ( A ) qPCR assay for osteoblast markers in cells 3 days after they were induced to differentiate. ( B , C ) ALP staining of WT and si Arhgap29 cells 7 days after they were induced to differentiate. ( D , E ) ARS staining of WT and si Arhgap29 cells after they were induced to differentiate for 14 days. ( F ) Quantitative analysis of alkaline phosphatase staining in cells ( n = 5). ( G ) Quantitative analysis of alizarin red staining in cells ( n = 5). All data: *** p < 0.001.

Article Snippet: After dewaxing and rehydration, the sections were incubated with the Arhgap29 primary antibody (sc-377022, Santa Cruz Biotechnology, Dallas, TX, USA) overnight at 4 °C, followed by incubation with a secondary antibody (Bioss, Beijing, China) for 30 min at 37 °C.

Techniques: In Vitro, Staining

Journal: Cancer Genomics & Proteomics

Article Title: ARHGAP29 Is Involved in Increased Invasiveness of Tamoxifen-resistant Breast Cancer Cells and its Expression Levels Correlate With Clinical Tumor Parameters of Breast Cancer Patients

doi: 10.21873/cgp.20454

Figure Lengend Snippet: Information on tissue of origin, pathology, grading, staging, TNM classification and assessment of ARHGAP29 expression. (–) not expressed, (+) slightly expressed, (++) moderately expressed and (+++) strongly expressed.

Article Snippet: Primary antibodies against ARHGAP29 1:2000 (#NBP1-05989, Novus Biologicals, Centennial, CO, USA), RhoC 1:2,000 (#GTX100546, GeneTex, Irvine, CA, USA), pAKT1 1:1,000 (#9271, Cell Signaling, Danvers, MA, USA), and GAPDH 1:2,000 (#5174S, Cell Signaling) were used.

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: MicroRNA-1291 promotes endometrial fibrosis by regulating the ArhGAP29-RhoA/ROCK1 signaling pathway in a murine model

doi: 10.3892/mmr.2017.7210

Figure Lengend Snippet: Immunohistochemical staining of ArhGAP29-RhoA/ROCK1 in human endometrium. ArhGAP29 staining in normal human endometrium was more obvious compared with endometrial tissue with severe IUAs; whereas, immunohistochemical staining of RhoA/ROCK1 in human endometrium with severe IUAs was more obvious compared with normal endometrial tissue. Scale bar, 50 µm. IUAs, intrauterine adhesions; ArhGAP29, Rho GTPase activating protein 29; ROCK1, Rho-associated coiled-coil containing protein kinase 1.

Article Snippet: Immunohistochemistry, using a commercial primary anti-ArhGAP29 antibody (NBP1-05989; Novus Biologicals Canada ULC, Oakville, ON, Canada), RhoA (sc179; 1:50; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and ROCK1 (ab45171; 1:200; Abcam, Cambridge, MA, USA) were used to investigate the differential expression of ArhGAP29 and RhoA/ROCK1 in severe IUAs compared with normal endometrial tissue.

Techniques: Immunohistochemical staining, Staining

Journal: Molecular Medicine Reports

Article Title: MicroRNA-1291 promotes endometrial fibrosis by regulating the ArhGAP29-RhoA/ROCK1 signaling pathway in a murine model

doi: 10.3892/mmr.2017.7210

Figure Lengend Snippet: Effect of miR-1291 antagomir on ArhGAP29-RhoA/ROCK1 pathway in murine uterine tissue. Immunofluorescence staining for ArhGAP29, RhoA and ROCK1 (green) and nuclear DNA (DAPI, blue) in murine uterine tissue from the IUAs group, miR-1291 antagomir group and the negative control group. Scale bar, 50 µm. ArhGAP29, Rho GTPase activating protein 29; ROCK1, Rho-associated coiled-coil containing protein kinase 1; IUAs, intrauterine adhesions; miR, microRNA.

Article Snippet: Immunohistochemistry, using a commercial primary anti-ArhGAP29 antibody (NBP1-05989; Novus Biologicals Canada ULC, Oakville, ON, Canada), RhoA (sc179; 1:50; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and ROCK1 (ab45171; 1:200; Abcam, Cambridge, MA, USA) were used to investigate the differential expression of ArhGAP29 and RhoA/ROCK1 in severe IUAs compared with normal endometrial tissue.

Techniques: Immunofluorescence, Staining, Negative Control

Journal: Molecular Medicine Reports

Article Title: MicroRNA-1291 promotes endometrial fibrosis by regulating the ArhGAP29-RhoA/ROCK1 signaling pathway in a murine model

doi: 10.3892/mmr.2017.7210

Figure Lengend Snippet: Effect of miR-1291 antagomir on ArhGAP29-RhoA/ROCK1 expression. (A) The expression levels of ArhGAP29, RhoA and ROCK1 were examined by reverse transcription-quantitative polymerase chain reaction. (B) Representative western-blot images and (C) densitometry of expression of ArhGAP29, RhoA and ROCK1 following western blotting analysis. ★ P<0.05, ★★ P<0.01 vs. the IUAs group; **P<0.01, ***P<0.001 vs. miR-1291 antagomir group. ArhGAP29, Rho GTPase activating protein 29; ROCK1, Rho-associated coiled-coil containing protein kinase 1; IUAs, intrauterine adhesions; miR, microRNA.

Article Snippet: Immunohistochemistry, using a commercial primary anti-ArhGAP29 antibody (NBP1-05989; Novus Biologicals Canada ULC, Oakville, ON, Canada), RhoA (sc179; 1:50; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and ROCK1 (ab45171; 1:200; Abcam, Cambridge, MA, USA) were used to investigate the differential expression of ArhGAP29 and RhoA/ROCK1 in severe IUAs compared with normal endometrial tissue.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot