polyclonal primary antibodies against cmip  (Proteintech)

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Correlation of CMIP expression with clinicopathological parameters from gastric cancer patients.

Article Snippet: Following blocking, the membranes were incubated with polyclonal primary antibodies against CMIP (cat. no. 12851-1-AP) and β-actin (cat. no. 20536-1-AP; both diluted at 1:1,000; ProteinTech Group, Inc., Chicago, IL, USA) overnight at 4°C.

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Expression of CMIP in gastric cancer tissues. (A) Immunohistochemistry analysis of CMIP in human gastric cancer tissues and normal gastric tissues. Human testes tissue and gastric cancer tissues were also examined as the CMIP positive and isotype negative controls, respectively. Representative images are presented (magnification, ×200). (B) Association between CMIP protein expression and relapse-free survival and overall survival. CMIP, c-Maf inducing protein.

Article Snippet: Following blocking, the membranes were incubated with polyclonal primary antibodies against CMIP (cat. no. 12851-1-AP) and β-actin (cat. no. 20536-1-AP; both diluted at 1:1,000; ProteinTech Group, Inc., Chicago, IL, USA) overnight at 4°C.

Techniques: Expressing, Immunohistochemistry

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Expression of CMIP in gastric cancer and normal tissues.

Article Snippet: Following blocking, the membranes were incubated with polyclonal primary antibodies against CMIP (cat. no. 12851-1-AP) and β-actin (cat. no. 20536-1-AP; both diluted at 1:1,000; ProteinTech Group, Inc., Chicago, IL, USA) overnight at 4°C.

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Knockdown of CMIP reduces the proliferation and enhances the apoptosis of MKN-28 cells. (A) The efficiency of CMIP knockdown by CMIP-siRNA was assessed by western blot analysis. Proliferation of MKN-28 cells transfected with CMIP-siRNA was measured using (B) cell counting (where Day 1=1×10 4 cells), (C) CCK-8 and (D) colony formation assays. (E) Apoptosis analysis of MKN-28 cells transfected with CMIP-siRNA was measured by flow cytometry using double-staining with Annexin V-fluorescein isothiocyanate and PI. CMIP, c-Maf inducing protein; si, small interfering; PI, propidium iodide; NC, negative control; OD, optical density. Data are presented as the mean ± standard error of the mean of at least 3 independent experiments. *P<0.05 and **P<0.01 vs. siNC.

Article Snippet: Following blocking, the membranes were incubated with polyclonal primary antibodies against CMIP (cat. no. 12851-1-AP) and β-actin (cat. no. 20536-1-AP; both diluted at 1:1,000; ProteinTech Group, Inc., Chicago, IL, USA) overnight at 4°C.

Techniques: Knockdown, Western Blot, Transfection, Cell Counting, CCK-8 Assay, Flow Cytometry, Double Staining, Negative Control

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Knockdown of CMIP suppresses the migration and invasion of MKN-28 cells. MKN-28 cells were transfected with the indicated siRNAs and the (A) migratory and (B) invasive abilities of the cells were evaluated (n=3; magnification, ×100). (C) A scratch was made in a confluent, adherent layer of MKN-28 cells that had undergone treatment with siNC or two different siRNAs against CMIP. Cells that had migrated into the wound were counted after 24 h (n=3). Representative images of the wounds are presented. Data are presented as the mean ± standard error of the mean of at least 3 independent experiments. *P<0.05 vs. siNC. CMIP, c-Maf inducing protein; si, small interfering; NC, negative control.

Article Snippet: Following blocking, the membranes were incubated with polyclonal primary antibodies against CMIP (cat. no. 12851-1-AP) and β-actin (cat. no. 20536-1-AP; both diluted at 1:1,000; ProteinTech Group, Inc., Chicago, IL, USA) overnight at 4°C.

Techniques: Knockdown, Migration, Transfection, Negative Control

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: CMIP is a direct target of miR-101-3p and CMIP regulates the expression of MDM2 and MAPK. (A) The TargetScan-predicted binding site between miR-101-3p and the 3′-UTR of CMIP. The mutant 3′-UTR of CMIP is also presented. (B) miR-101-3p was down-regulated in gastric cancer tissues compared with normal gastric tissues. *P<0.05 vs. Normal. (C) Luciferase assay of MKN-28 cells cotransfected with miR-101-3p mimic or NC, and a luciferase reporter containing CMIP 3′-UTR wildtype or mutant constructs. *P<0.05 vs. NC. (D) MKN-28 cells were transfected with miR-101-3p mimics or NC. miR-101-3p overexpression inhibited the protein expression of CMIP. β-actin served as a loading control. (E) Cells transfected with CMIP-siRNA demonstrated a significant decrease in MDM2 and MAPK mRNA expression compared with cells transfected with siNC. All data were presented as the mean ± standard error of the mean of at least 3 independent experiments. *P<0.05 vs. siNC. CMIP, c-Maf inducing protein; MAPK, mitogen activated protein kinase; miR, microRNA; NC, negative control; si, small interfering; 3′UTR, 3-untranslated region; 3′UTR-WT, wild type 3′UTR construct; 3′UTR-MUT, mutant 3′UTR construct; MAPK, mitogen activated protein kinase.

Article Snippet: Following blocking, the membranes were incubated with polyclonal primary antibodies against CMIP (cat. no. 12851-1-AP) and β-actin (cat. no. 20536-1-AP; both diluted at 1:1,000; ProteinTech Group, Inc., Chicago, IL, USA) overnight at 4°C.

Techniques: Expressing, Binding Assay, Mutagenesis, Luciferase, Construct, Transfection, Over Expression, Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Correlation of CMIP expression with clinicopathological parameters from gastric cancer patients.

Article Snippet: Immunohistochemistry (IHC) analyses of CMIP protein expression was performed using a Two-Step Histostaining kit (Fuzhou Maixin Biotech Co., Ltd., Fuzhou, China) with a polyclonal primary antibody against CMIP (1:100; cat. no. 12851-1-AP; Proteintech Group, Inc., Chicago, USA).

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Expression of CMIP in gastric cancer tissues. (A) Immunohistochemistry analysis of CMIP in human gastric cancer tissues and normal gastric tissues. Human testes tissue and gastric cancer tissues were also examined as the CMIP positive and isotype negative controls, respectively. Representative images are presented (magnification, ×200). (B) Association between CMIP protein expression and relapse-free survival and overall survival. CMIP, c-Maf inducing protein.

Article Snippet: Immunohistochemistry (IHC) analyses of CMIP protein expression was performed using a Two-Step Histostaining kit (Fuzhou Maixin Biotech Co., Ltd., Fuzhou, China) with a polyclonal primary antibody against CMIP (1:100; cat. no. 12851-1-AP; Proteintech Group, Inc., Chicago, USA).

Techniques: Expressing, Immunohistochemistry

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Expression of CMIP in gastric cancer and normal tissues.

Article Snippet: Immunohistochemistry (IHC) analyses of CMIP protein expression was performed using a Two-Step Histostaining kit (Fuzhou Maixin Biotech Co., Ltd., Fuzhou, China) with a polyclonal primary antibody against CMIP (1:100; cat. no. 12851-1-AP; Proteintech Group, Inc., Chicago, USA).

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Knockdown of CMIP reduces the proliferation and enhances the apoptosis of MKN-28 cells. (A) The efficiency of CMIP knockdown by CMIP-siRNA was assessed by western blot analysis. Proliferation of MKN-28 cells transfected with CMIP-siRNA was measured using (B) cell counting (where Day 1=1×10 4 cells), (C) CCK-8 and (D) colony formation assays. (E) Apoptosis analysis of MKN-28 cells transfected with CMIP-siRNA was measured by flow cytometry using double-staining with Annexin V-fluorescein isothiocyanate and PI. CMIP, c-Maf inducing protein; si, small interfering; PI, propidium iodide; NC, negative control; OD, optical density. Data are presented as the mean ± standard error of the mean of at least 3 independent experiments. *P<0.05 and **P<0.01 vs. siNC.

Article Snippet: Immunohistochemistry (IHC) analyses of CMIP protein expression was performed using a Two-Step Histostaining kit (Fuzhou Maixin Biotech Co., Ltd., Fuzhou, China) with a polyclonal primary antibody against CMIP (1:100; cat. no. 12851-1-AP; Proteintech Group, Inc., Chicago, USA).

Techniques: Knockdown, Western Blot, Transfection, Cell Counting, CCK-8 Assay, Flow Cytometry, Double Staining, Negative Control

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: Knockdown of CMIP suppresses the migration and invasion of MKN-28 cells. MKN-28 cells were transfected with the indicated siRNAs and the (A) migratory and (B) invasive abilities of the cells were evaluated (n=3; magnification, ×100). (C) A scratch was made in a confluent, adherent layer of MKN-28 cells that had undergone treatment with siNC or two different siRNAs against CMIP. Cells that had migrated into the wound were counted after 24 h (n=3). Representative images of the wounds are presented. Data are presented as the mean ± standard error of the mean of at least 3 independent experiments. *P<0.05 vs. siNC. CMIP, c-Maf inducing protein; si, small interfering; NC, negative control.

Article Snippet: Immunohistochemistry (IHC) analyses of CMIP protein expression was performed using a Two-Step Histostaining kit (Fuzhou Maixin Biotech Co., Ltd., Fuzhou, China) with a polyclonal primary antibody against CMIP (1:100; cat. no. 12851-1-AP; Proteintech Group, Inc., Chicago, USA).

Techniques: Knockdown, Migration, Transfection, Negative Control

Journal: Molecular Medicine Reports

Article Title: CMIP is oncogenic in human gastric cancer cells

doi: 10.3892/mmr.2017.7541

Figure Lengend Snippet: CMIP is a direct target of miR-101-3p and CMIP regulates the expression of MDM2 and MAPK. (A) The TargetScan-predicted binding site between miR-101-3p and the 3′-UTR of CMIP. The mutant 3′-UTR of CMIP is also presented. (B) miR-101-3p was down-regulated in gastric cancer tissues compared with normal gastric tissues. *P<0.05 vs. Normal. (C) Luciferase assay of MKN-28 cells cotransfected with miR-101-3p mimic or NC, and a luciferase reporter containing CMIP 3′-UTR wildtype or mutant constructs. *P<0.05 vs. NC. (D) MKN-28 cells were transfected with miR-101-3p mimics or NC. miR-101-3p overexpression inhibited the protein expression of CMIP. β-actin served as a loading control. (E) Cells transfected with CMIP-siRNA demonstrated a significant decrease in MDM2 and MAPK mRNA expression compared with cells transfected with siNC. All data were presented as the mean ± standard error of the mean of at least 3 independent experiments. *P<0.05 vs. siNC. CMIP, c-Maf inducing protein; MAPK, mitogen activated protein kinase; miR, microRNA; NC, negative control; si, small interfering; 3′UTR, 3-untranslated region; 3′UTR-WT, wild type 3′UTR construct; 3′UTR-MUT, mutant 3′UTR construct; MAPK, mitogen activated protein kinase.

Article Snippet: Immunohistochemistry (IHC) analyses of CMIP protein expression was performed using a Two-Step Histostaining kit (Fuzhou Maixin Biotech Co., Ltd., Fuzhou, China) with a polyclonal primary antibody against CMIP (1:100; cat. no. 12851-1-AP; Proteintech Group, Inc., Chicago, USA).

Techniques: Expressing, Binding Assay, Mutagenesis, Luciferase, Construct, Transfection, Over Expression, Control, Negative Control