primary antibodies against trpm8  (Alomone Labs)

Journal: Cancer medicine

Article Title: TRPM8 overexpression suppresses hepatocellular carcinoma progression and improves survival by modulating the RTP3/STAT3 pathway.

doi: 10.1002/cam4.70109

Figure Lengend Snippet: FIGURE 4 TRPM8 level determination and TRPM8-silencing cell function experiments. (A–D) TRPM8 inhibition in PLC/PRF/5 and HUH-7 cells transfected with shTRPM8 lentivirus was verified by Western blot and qRT-PCR. (E–I) Effect of TRPM8 loss-of-function on PLC/RPF/5 and HUH-7 cell proliferation was determined by colony formation assay (E), CCK-8 (F-G) and EdU assay (H, I). (J, K) The effects of TRPM8 inhibition on the cell migration and invasion of PLC/PRF/5 and HUH-7 cells were detected using Transwell assays. Scale bars: 100 μm. *p < 0.05.

Article Snippet: Primary antibodies against TRPM8 (Zen- bioscience, Chengdu, China), RTP3 (Epigentek, USA), STAT3 (Proteintech, Wuhan, China) and GAPDH (Proteintech, Wuhan, China) were employed.

Techniques: Cell Function Assay, Inhibition, Transfection, Western Blot, Quantitative RT-PCR, Colony Assay, CCK-8 Assay, EdU Assay, Migration

Journal: Cancer medicine

Article Title: TRPM8 overexpression suppresses hepatocellular carcinoma progression and improves survival by modulating the RTP3/STAT3 pathway.

doi: 10.1002/cam4.70109

Figure Lengend Snippet: FIGURE 6 AD80 promoted the expression of TRPM8 and inhibited the proliferation, migration and invasion of HCC cells in vitro. (A, B) Calcium imaging was used to detected the changes of the fluorescence intensity of WS12 (A) and AD80 (B) groups on time in SNU-449 cells. (C, D) Effects of WS12 (C) and AD80 (D) on the expression of TRPM8 in SNU-449 cells were detected by Western blot. (E-G) EdU (E) and CCK-8 assays (F) were used to detect the effect of WS12 on the proliferation of SNU-449. (G) The effects of WS12 on the cell migration and invasion of SSNU-449 cells were detected using Transwell assays. (H–J) EdU (H) and CCK-8 assays (I) were used to detect the effect of WS12 on the proliferation of SSNU-449. (J) The effects of AD80 on the cell migration and invasion of SNU-449 cells were detected using Transwell assays. Scale bars: 100 μm. *p < 0.05.

Article Snippet: Primary antibodies against TRPM8 (Zen- bioscience, Chengdu, China), RTP3 (Epigentek, USA), STAT3 (Proteintech, Wuhan, China) and GAPDH (Proteintech, Wuhan, China) were employed.

Techniques: Expressing, Migration, In Vitro, Imaging, Fluorescence, Western Blot, CCK-8 Assay

Journal: Cancer medicine

Article Title: TRPM8 overexpression suppresses hepatocellular carcinoma progression and improves survival by modulating the RTP3/STAT3 pathway.

doi: 10.1002/cam4.70109

Figure Lengend Snippet: FIGURE 7 Schematic depicting the possible mechanism of the TRPM8- mediated RTP3/STAT3 pathway in HCC.

Article Snippet: Primary antibodies against TRPM8 (Zen- bioscience, Chengdu, China), RTP3 (Epigentek, USA), STAT3 (Proteintech, Wuhan, China) and GAPDH (Proteintech, Wuhan, China) were employed.

Techniques:

Journal: PLoS ONE

Article Title: TRPM8 Channel Activation Induced by Monoterpenoid Rotundifolone Underlies Mesenteric Artery Relaxation

doi: 10.1371/journal.pone.0143171

Figure Lengend Snippet: (A) TRPM8 mRNA expression by semi-quantitative PCR in superior mesenteric artery with endothelium removed (n = 3). C, control; P1, 1 μL prostate RT reaction; P2, 2 μL prostate RT reaction, Me, 2 μL mesenteric artery RT reaction. (B) Western blot analysis of TRPM8 protein expression in rat vascular tissue. Molecular weight of TRPM8 was 125 KDa. Bottom panel: β actin used to demonstrate protein loading (at 42kDa).

Article Snippet: The membrane was blocked with 5% (wt/vol) milk in TBS containing 0.1% Tween 20 (TBST) for 1 h at room temperature, followed by overnight incubation at 4°C with the specific primary rabbit polyclonal antibody against TRPM8 (1:1000 dilution; Abcam, Cambridge, Massachusetts, United States).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Western Blot, Molecular Weight

Journal: Oncology Letters

Article Title: Overexpression of short TRPM8 variant α promotes cell migration and invasion, and decreases starvation-induced apoptosis in prostate cancer LNCaP cells

doi: 10.3892/ol.2015.3373

Figure Lengend Snippet: (A) Western blot analysis indicating that overexpression sM8α-His fusion protein reduced the expression of p-JNK and increased the expression of active MMP-2 (64 kDa) in LNCaP cells. GAPDH was used as the inner control. (B) sM8α-His fusion protein was significantly overexpressed in LNCaP-sM8α cells. (C) The expression of p-JNK protein was significantly reduced in LNCaP-sM8α cells compared with LNCaP and LNCaP-NC cells. However, there were no significant changes in the expression of (D) p-ERK1/2 and (F) p-p38 or (E) full-length TRPM8 protein. (G) MMP-2 was activated, as indicated by upregulation of 64-kDa MMP-2 and downregulation of 72-kDa MMP-2 in LNCaP-sM8α cells. (H) There were no remarkable differences in the expression of MMP-9 in LNCaP, LNCaP-NC and LNCaP-sM8α cells. *P<0.05. NC, negative control; sM8α, short transient receptor potential melastatin 8α; TRPM8, transient receptor potential melastatin 8; p-ERK, phosphorylated-extracellular signal-regulated kinase 1/2; p-JNK, phosphorylated-c-Jun N-terminal kinase; MMP, matrix metalloproteinase; GAPDH, glyceraldehyde phosphate dehydrogenase.

Article Snippet: Primary antibodies against human TRPM8 (rabbit polyclonal; catalog no. ACC-049; Alomone Labs, Jerusalem, Israel; 1:500 dilution), MMP-2 (rabbit monoclonal IgG; catalog no. 13132; Cell Signaling Technology, Inc., Danvers, MA, USA; 1:1,000 dilution), MMP-9 (rabbit monoclonal IgG; catalog no. 13667, Cell Signaling Technology, Inc.; 1:1,000 dilution), GAPDH (rabbit polyclonal IgG; catalog no. sc-25778, Santa Cruz Biotechnology, Inc., Dallas, TX, USA; 1:1000 dilution), MAPK family [rabbit anti-p38, ERK1/2 (p44/42) and JNK (catalog no. 9926), and phospho (p-)p38, p-ERK1/2 and p-JNK (catalog no. 9910); Cell Signaling Technology, Inc.] and His-tag (mouse monoclonal IgG2a; catalog no. D291-3, Medical & Biological Laboratories Co., Ltd., Nagoya, Japan) were applied overnight at 4°C.

Techniques: Western Blot, Over Expression, Expressing, Negative Control