rabbit polyclonal antibody against trpm8  (Danaher Inc)

Journal: Cell & Bioscience

Article Title: The mechanism of Annexin A1 to modulate TRPV1 and nociception in dorsal root ganglion neurons

doi: 10.1186/s13578-021-00679-1

Figure Lengend Snippet: AnxA1 deletion does not alter the expression of FPR2, TRPV1 and other nociceptors. a Representative images of double immunofluorescent staining and b Venn diagram of the percentages of FPR2 positive and TRPV1 positive neurons in DRG sections from AnxA1 −/− mice and WT littermates labeled for FPR2 and TRPV1 as indicated. Scale bar, 100 μm. c Representative western blots band images and the quantification of the indicated proteins d ANXA1 ( AnxA1 −/− versus control group, P = 0.45, Student’s t-test. n = 6 in control group, n = 6 in AnxA1 −/− group), and e TRPV1 AnxA1 −/− versus control group, P = 0.32, Student’s t-test. n = 6 in WT group, n = 6 in AnxA1 −/− group) in DRG tissues between AnxA1 −/− mice and control littermates. f Representative band images and the quantification of other nociceptors such as g TRPA1 and h TRPM8 in L4-6 DRG tissues between AnxA1 −/− mice and control littermates ( AnxA1 −/− versus control, P = 0.82, P = 0.85, Student’s t-test. n = 6 in control group, n = 6 in AnxA1 −/− group). The relative density of the protein band image in control group was normalized to 1. All data are represented as mean ± SD

Article Snippet: For immunofluorescent staining, DRG sections were treated with 10% TritonX-100 for 30 min. Nonspecific binding sites were blocked with 5% fetal bovine serum for 30 min after washing with PBS for 10 min, followed by incubation with rabbit polyclonal antibody against TRPV1 (1:500, Cat#ab6166, Abcam), rabbit polyclonal antibody against TRPA1 (1:2000, Cat#OST00061W, Invitrogen), rabbit polyclonal antibody against TRPM8 (1:1000, Cat#OSR00077W, Invitrogen), mouse monoclonal antibody against FPR2 (1:1000, Cat#H00002358-M02, Abnova), and mouse monoclonal antibody against Calmodulin (1:1000, Cat#05-173, Sigma-Aldrich), overnight at 4 °C.

Techniques: Expressing, Staining, Labeling, Western Blot

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, Western Blot, Molecular Weight

Journal: The Korean Journal of Pain

Article Title: Effects of Nefopam on Streptozotocin-Induced Diabetic Neuropathic Pain in Rats

doi: 10.3344/kjp.2014.27.4.326

Figure Lengend Snippet: Effect of a nefopam pretreatment on the transient receptor potential ankyrin 1 (TRPA1) and melastatin 8 (TRPM8) protein expression levels in the dorsal root ganglion (DRG) of streptozotocin (STZ)-induced diabetic rats. Pretreatment with nefopam (30 mg/kg) was performed intraperitoneally 30 min prior to an intraperitoneal injection of STZ (60 mg/kg). To evaluate the expressions of TRPA1 (A) and TRPM8 (B) proteins located in the L4-L6 DRG, a Western blot analysis was performed 4 weeks after the final drug administration. The specific signals for TRPA1 and TRPM8 were quantified and plotted (lower panel). β-actin was used as an internal loading control. STZ injections did not significantly affect the basal level of the TRPA1 protein in the DRG, which was not altered by the nefopam pretreatment. The TRPM8 protein level in the DRG significantly increased 4 weeks after the STZ injection, which was reduced by the nefopam pretreatment. *** P < 0.001 compared to the control group. ††† P < 0.001 compared to the STZ group.

Article Snippet: After blocking with 5% goat serum, 0.3% Triton X-100, and 1% bovine serum albumin in phosphate-buffered saline (PBS-T), double-label immunofluorescence was conducted by incubating the paraffin sections overnight at 4℃ with rabbit polyclonal antibodies against TRPM8 (1:1000, Abcam), with the IgG fraction of the mouse polyclonal antibody against the N52 clone of neurofilament 200 (NF200, 1:200; Sigma-Aldrich), followed by incubation with Alexa Flour 555 goat anti-rabbit IgG (red; 1:500; Invitrogen, Carlsbad, CA, USA) and Alexa Flour 488 goat anti-mouse IgG (green; 1:500; Invitrogen) for 2 hours at 20℃.

Techniques: Expressing, Injection, Western Blot

Journal: The Korean Journal of Pain

Article Title: Effects of Nefopam on Streptozotocin-Induced Diabetic Neuropathic Pain in Rats

doi: 10.3344/kjp.2014.27.4.326

Figure Lengend Snippet: Typical double immunofluorescence staining of transient receptor potential melastatin 8 (TRPM8) and neurofilament 200 (NF200) in the rat dorsal root ganglion (DRG) of streptozotocin (STZ)-induced diabetic rats. Pretreatment with nefopam (30 mg/kg) was performed intraperitoneally 30 min prior to an intraperitoneal injection of STZ (60 mg/kg). Four weeks after the final drug treatment, L5 DRG samples were immunostained with TRPM8 and neurofilament 200 (NF200), a marker of myelinated neurons. The colocalization of TRPM8 was visualized in a merged image.

Article Snippet: After blocking with 5% goat serum, 0.3% Triton X-100, and 1% bovine serum albumin in phosphate-buffered saline (PBS-T), double-label immunofluorescence was conducted by incubating the paraffin sections overnight at 4℃ with rabbit polyclonal antibodies against TRPM8 (1:1000, Abcam), with the IgG fraction of the mouse polyclonal antibody against the N52 clone of neurofilament 200 (NF200, 1:200; Sigma-Aldrich), followed by incubation with Alexa Flour 555 goat anti-rabbit IgG (red; 1:500; Invitrogen, Carlsbad, CA, USA) and Alexa Flour 488 goat anti-mouse IgG (green; 1:500; Invitrogen) for 2 hours at 20℃.

Techniques: Double Immunofluorescence Staining, Injection, Marker