rabbit monoclonal anti mouse braf  (Cell Signaling Technology Inc)

Journal: Oncology Letters

Article Title: Analysis of the clonal origin and differences in the biological behavior of multifocal papillary thyroid carcinoma

doi: 10.3892/ol.2024.14677

Figure Lengend Snippet: Histology of individual multifocal papillary thyroid carcinoma samples. Representative images of (A) BRAF protein-positive, (B) BRAF protein-negative, (C) TERT protein-positive and (D) TERT protein-negative samples. Magnification, ×40. BRAF, v-raf murine sarcoma viral oncogene homolog B1; TERT, telomerase reverse transcriptase.

Article Snippet: The primary antibodies used were mouse anti-human BRAF monoclonal antibody (cat no. TA500431) and rabbit anti-human TERT polyclonal antibody (cat. no. TA324097), both from OriGene Technologies, Inc, a dilution of 1:200.

Techniques: Reverse Transcription

Journal: Oncology Letters

Article Title: Analysis of the clonal origin and differences in the biological behavior of multifocal papillary thyroid carcinoma

doi: 10.3892/ol.2024.14677

Figure Lengend Snippet: BRAF V600E mutation in multifocal papillary thyroid carcinoma. Representative (A) BRAF V600E mutant and (B) wild-type amplification plots generated using quantitative PCR. The red marking represents the threshold line. BRAF, v-raf murine sarcoma viral oncogene homolog B1; BRAF-IC, BRAF internal reference gene detection curve; BRAF-W, BRAF V600E wild-type gene detection curve; BRAF-M, BRAF V600E mutation gene detection curve; DRn, change in relative fluorescence normalized.

Article Snippet: The primary antibodies used were mouse anti-human BRAF monoclonal antibody (cat no. TA500431) and rabbit anti-human TERT polyclonal antibody (cat. no. TA324097), both from OriGene Technologies, Inc, a dilution of 1:200.

Techniques: Mutagenesis, Amplification, Generated, Real-time Polymerase Chain Reaction, Fluorescence

Journal: Oncology Letters

Article Title: Analysis of the clonal origin and differences in the biological behavior of multifocal papillary thyroid carcinoma

doi: 10.3892/ol.2024.14677

Figure Lengend Snippet: Gene mutation and protein expression in cases of multifocal papillary thyroid carcinoma with different clonal origins.

Article Snippet: The primary antibodies used were mouse anti-human BRAF monoclonal antibody (cat no. TA500431) and rabbit anti-human TERT polyclonal antibody (cat. no. TA324097), both from OriGene Technologies, Inc, a dilution of 1:200.

Techniques: Mutagenesis, Expressing

Journal: Oncology Letters

Article Title: Analysis of the clonal origin and differences in the biological behavior of multifocal papillary thyroid carcinoma

doi: 10.3892/ol.2024.14677

Figure Lengend Snippet: Figure 1. Histology of individual multifocal papillary thyroid carcinoma samples. Representative images of (A) BRAF protein‑positive, (B) BRAF protein‑ negative, (C) TERT protein‑positive and (D) TERT protein‑negative samples. Magnification, x40. BRAF, v‑raf murine sarcoma viral oncogene homolog B1; TERT, telomerase reverse transcriptase.

Article Snippet: The primary antibodies used were mouse anti‐human BRAF monoclonal antibody (cat no. TA500431) and rabbit anti‐human TERT polyclonal antibody (cat. no. TA324097), both from OriGene Technologies, Inc, a dilution of 1:200.

Techniques: Reverse Transcription

Journal: Oncology Letters

Article Title: Analysis of the clonal origin and differences in the biological behavior of multifocal papillary thyroid carcinoma

doi: 10.3892/ol.2024.14677

Figure Lengend Snippet: Figure 2. BRAF V600E mutation in multifocal papillary thyroid carcinoma. Representative (A) BRAF V600E mutant and (B) wild‑type amplification plots generated using quantitative PCR. The red marking represents the threshold line. BRAF, v‑raf murine sarcoma viral oncogene homolog B1; BRAF‑IC, BRAF internal reference gene detection curve; BRAF‑W, BRAF V600E wild‑type gene detection curve; BRAF‑M, BRAF V600E mutation gene detection curve; DRn, change in relative fluorescence normalized.

Article Snippet: The primary antibodies used were mouse anti‐human BRAF monoclonal antibody (cat no. TA500431) and rabbit anti‐human TERT polyclonal antibody (cat. no. TA324097), both from OriGene Technologies, Inc, a dilution of 1:200.

Techniques: Mutagenesis, Amplification, Generated, Real-time Polymerase Chain Reaction, Fluorescence

Journal: Journal of translational medicine

Article Title: BRAF D594A mutation defines a unique biological and immuno-modulatory subgroup associated with functional CD8 + T cell infiltration in colorectal cancer.

doi: 10.1186/s12967-023-04606-5

Figure Lengend Snippet: Fig. 1 D594A mutation confers lower aggressiveness on MC38 cells. A Immunoblotting of BRAF level in WT and mutant MC38 cells. Actin serves as an internal reference. B Representative immunoblotting of MEK, pMEK, ERK1/2, pERK1/2 and Cyclin D1 in the starvation group (right) and stimulation group (left). Cells were exposed to EGF (100 ng/mL) for 15 min after starvation for 36 h in stimulation group. C Proliferation ability of BRAF WT and mutant MC38 cells at 24 h, 48 h and 72 h detected by CCK-8 assay. D Detection of apoptosis by annexin V/PI staining after induction of FCCP for 36 h. E Growth ability of BRAF WT and mutant MC38 cells analyzed by clone formation assay. F, G Migration and invasion abilities of BRAF WT and mutant MC38 cells analyzed by transwell assays, respectively. Three independent replicates were performed for above experiments. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001

Article Snippet: Rabbit monoclonal anti-mouse BRAF, THBS1, p-ERK, p-MEK, ERK, MEK and Cyclin D1 (Cat Nr. 14814, 37879, 4370, 9154, 4695, 4694 and 2978, Cell Signaling Technology, Germany), rabbit monoclonal anti-rabbit β-actin (Cat Nr. AC026, ABclonal, USA) were used as primary antibodies in the study.

Techniques: Mutagenesis, Western Blot, CCK-8 Assay, Staining, Tube Formation Assay, Migration

Journal: Journal of translational medicine

Article Title: BRAF D594A mutation defines a unique biological and immuno-modulatory subgroup associated with functional CD8 + T cell infiltration in colorectal cancer.

doi: 10.1186/s12967-023-04606-5

Figure Lengend Snippet: Fig. 2 BRAF mutation modulates the expression of immune regulatory molecules. A The effect of IFN-γ treatment on the survival of tumor cells analyzed by flow cytometry. B, C The basal and induced expressions of PD-L1 in BRAF WT and mutant MC38 cells detected by flow cytometry and qRT-PCR. D The basal and induced expressions of MHC class I in BRAF WT and mutant cells detected by flow cytometry. E–G The basal and induced expressions of Psmb8, H2-D1 and B2M in BRAF WT and mutant cells detected by qRT-PCR. Cells were treated by 20 ng/mL IFN-γ for 48 h. Three independent replicates were performed for above experiments. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001

Article Snippet: Rabbit monoclonal anti-mouse BRAF, THBS1, p-ERK, p-MEK, ERK, MEK and Cyclin D1 (Cat Nr. 14814, 37879, 4370, 9154, 4695, 4694 and 2978, Cell Signaling Technology, Germany), rabbit monoclonal anti-rabbit β-actin (Cat Nr. AC026, ABclonal, USA) were used as primary antibodies in the study.

Techniques: Mutagenesis, Expressing, Flow Cytometry, Quantitative RT-PCR

Journal: Journal of translational medicine

Article Title: BRAF D594A mutation defines a unique biological and immuno-modulatory subgroup associated with functional CD8 + T cell infiltration in colorectal cancer.

doi: 10.1186/s12967-023-04606-5

Figure Lengend Snippet: Fig. 3 BRAF mutation influences the panel of TILs in transplanted tumor. BRAF WT and mutant MC38 cells were inoculated subcutaneously in homologous C57BL/6J mice (n = 5 mice per group). The tumor sizes, weights and growth curves of the four groups were shown as (A–C), respectively. D Analysis of the population of TILs by flow cytometry, including CD3, CD4, CD8, NK cells and Macrophage. E Representative immunohistochemical staining for CD8α in paraffin section of transplanted tumors (magnification 400 ×). *P < 0.05; **P < 0.01; ***P < 0.001

Article Snippet: Rabbit monoclonal anti-mouse BRAF, THBS1, p-ERK, p-MEK, ERK, MEK and Cyclin D1 (Cat Nr. 14814, 37879, 4370, 9154, 4695, 4694 and 2978, Cell Signaling Technology, Germany), rabbit monoclonal anti-rabbit β-actin (Cat Nr. AC026, ABclonal, USA) were used as primary antibodies in the study.

Techniques: Mutagenesis, Flow Cytometry, Immunohistochemical staining, Staining, Paraffin Section

Journal: Journal of translational medicine

Article Title: BRAF D594A mutation defines a unique biological and immuno-modulatory subgroup associated with functional CD8 + T cell infiltration in colorectal cancer.

doi: 10.1186/s12967-023-04606-5

Figure Lengend Snippet: Fig. 5 D594A mutation reprograms transcriptome involved in cell chemotaxis and upregulates THBS1 expression via ATF3. A Analysis of differentially expressed genes (DEGs) between D594A and WT cells by RNA-seq. B Functional enrichment of DEGs by online Metascape analysis. C The potential relationship between THBS1 and CXCL10 revealed by bioinformation prediction. D The basal and induced expressions of THBS1 in BRAF WT and mutant cells detected by qRT-PCR. Cells were treated by 20 ng/mL IFN-γ for 48 h. E The protein level of THBS1 in BRAF WT and mutant cells detected by western blot. F The putative binding sites of ATF3 on THBS1 promoter predicted by jaspar website. G The ATF3 mRNA level in BRAF WT and mutant cells detected by qRT-PCR. H The transcriptional regulation of THBS1 by ATF3 examed by dual-luciferase reporter assay. EV, empty vector; NC, negative control. *P < 0.05; **P < 0.01; ***P < 0.001

Article Snippet: Rabbit monoclonal anti-mouse BRAF, THBS1, p-ERK, p-MEK, ERK, MEK and Cyclin D1 (Cat Nr. 14814, 37879, 4370, 9154, 4695, 4694 and 2978, Cell Signaling Technology, Germany), rabbit monoclonal anti-rabbit β-actin (Cat Nr. AC026, ABclonal, USA) were used as primary antibodies in the study.

Techniques: Mutagenesis, Chemotaxis Assay, Expressing, RNA Sequencing, Functional Assay, Quantitative RT-PCR, Western Blot, Binding Assay, Luciferase, Reporter Assay, Plasmid Preparation, Negative Control

Journal: Journal of translational medicine

Article Title: BRAF D594A mutation defines a unique biological and immuno-modulatory subgroup associated with functional CD8 + T cell infiltration in colorectal cancer.

doi: 10.1186/s12967-023-04606-5

Figure Lengend Snippet: Fig. 6 THBS1 improves CXCL9/CXCL10 chemokines release in D594A mutant cells. A The mRNA levels of CXCL9, CXCL10 and CXCL11 in BRAF WT and mutant MC38 cells detected by qRT-PCR. B The concentrations of CXCL9 and CXCL10 in the supernatant of BRAF WT and mutant cells analyzed by ELISA. C The concentrations of CXCL9 and CXCL10 in the supernatant of hTHBS1 and hctrl cells detected by ELISA. D The mRNA expressions of CXCL9 and CXCL10 in hTHBS1 and hctrl cells detected by qRT-PCR. E The concentrations of CXCL9 and CXCL10 in the supernatant of siTHBS1 and sictrl cells analyzed by ELISA. F The mRNA expressions of CXCL9 and CXCL10 in siTHBS1 and sictrl cells detected by qRT-PCR. Cells were treated by 20 ng/mL IFN-γ for 48 h. Three independent replicates were performed for above experiments. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001

Article Snippet: Rabbit monoclonal anti-mouse BRAF, THBS1, p-ERK, p-MEK, ERK, MEK and Cyclin D1 (Cat Nr. 14814, 37879, 4370, 9154, 4695, 4694 and 2978, Cell Signaling Technology, Germany), rabbit monoclonal anti-rabbit β-actin (Cat Nr. AC026, ABclonal, USA) were used as primary antibodies in the study.

Techniques: Mutagenesis, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay