rabbit polyclonal antibody trpm8 specific  (Alomone Labs)

Journal: Frontiers in Cell and Developmental Biology

Article Title: TRPM8 Channel Promotes the Osteogenic Differentiation in Human Bone Marrow Mesenchymal Stem Cells

doi: 10.3389/fcell.2021.592946

Figure Lengend Snippet: TRPM8 channel expression in hBM-MSCs. (A) RT-PCR from total hBM-MSCs RNA. cDNA fragment corresponding to TRPM8 transcript (144 bp) in hBM-MSCs is shown. As controls we used HEK cells transfected with TRPM8 and non-transfected cells, respectively. RPL 27 (Name of the Ribosomal proteinL27, 120 bp) is the housekeeping gene ( n = 3). (B) Western blot detected TRPM8 protein in hBM-MSCs and cells transfected with TRPM8 (130 kDa). β-actin (45kDA) was used as a load control. As a negative control we used untransfected cells. (C) Percentage of TRPM8 total and membrane expression in hBM-MSCs and control cells transfection ( n = 3). (D) TRPM8 expression pattern in permeabilized hBM-MSCs (left panel) and control of unspecific secondary antibody binding (right panel) ( n = 4). The images presented in the figure shown confocal images of Alexa fluor 488 (TRPM8, green) and DAPI (blue).

Article Snippet: Cells were washed with TBS, incubated with TBS-BSA 2% to block non-specific binding and left overnight with 1/100 of rabbit polyclonal antibody TRPM8 specific (ACC-049, Alomone Labs).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Western Blot, Negative Control, Binding Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: TRPM8 Channel Promotes the Osteogenic Differentiation in Human Bone Marrow Mesenchymal Stem Cells

doi: 10.3389/fcell.2021.592946

Figure Lengend Snippet: TRPM8 hBM-MSC localization. Maximum image volume projection of hBM-MSCs after double staining with the antibody TRPM8 conjugated with Alexa 488 and Mitotracker Red FM (colocalization index 0.03) (Overlap 0.36) (A) , Lisotracker Red (colocalization index 0.02) (overlap 0.49) (B) , and Alexa 647 calnexin (colocalization index 0.14) (Overlap 0.60) (C) . (D) Selected area for the colocalization analysis (perinuclear region). (E) Colocalization analysis ( n = 3) experiments.

Article Snippet: Cells were washed with TBS, incubated with TBS-BSA 2% to block non-specific binding and left overnight with 1/100 of rabbit polyclonal antibody TRPM8 specific (ACC-049, Alomone Labs).

Techniques: Double Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: TRPM8 Channel Promotes the Osteogenic Differentiation in Human Bone Marrow Mesenchymal Stem Cells

doi: 10.3389/fcell.2021.592946

Figure Lengend Snippet: hBM-MSCs express functional TRPM8 channels. (A,B) Currents evoked in response to a voltage pulse protocol. Current- voltage (IV) curves in whole cell configuration in hBM-MSCs cells before (control) and after the addition of menthol 500 μM and BCTC 10 μM ( n : 4). (Insert: Voltage step protocol). TRPM8 currents were calculated by subtracting the residual current after the addition of BCTC current from the current activated by menthol. (B) IV in response to a voltage ramp protocol in configuration of whole cell before (control, continues line) and after the addition of menthol 500 μM (dashed line) and BCTC 10 μM (dotted line) ( n = 4). The ramp protocol used for the acquisition of the records is indicated in the insert. (C) Current density (pA/pF) from hBM-MSC at −80 mV and +80 mV, before and after menthol addition. * P < 0.05. Data are mean ± SD ( n = 3).

Article Snippet: Cells were washed with TBS, incubated with TBS-BSA 2% to block non-specific binding and left overnight with 1/100 of rabbit polyclonal antibody TRPM8 specific (ACC-049, Alomone Labs).

Techniques: Functional Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: TRPM8 Channel Promotes the Osteogenic Differentiation in Human Bone Marrow Mesenchymal Stem Cells

doi: 10.3389/fcell.2021.592946

Figure Lengend Snippet: Osteogenic differentiation in hBM-MSCs cells is regulated by exposure to TRPM8 channel modulators. (A) Percentage of differentiated area from hBM-MSCs, detected with Alizarin Red (n = 3). Negative control (Control -), **** P < 0.0001; 100 μM menthol, ** P < 0.001; 20 μM BCTC, * P < 0.01; 10 μM Icilin, ** P < 0.001; (n = 3). (B) Rate of change measured by qPCR of the ALPL gene after 14 days of osteogenic differentiation; Negative control (Control -), * P < 0.01; BCTC 20 μM, ** P < 0.001; 100 μM Menthol, ** P < 0.001.

Article Snippet: Cells were washed with TBS, incubated with TBS-BSA 2% to block non-specific binding and left overnight with 1/100 of rabbit polyclonal antibody TRPM8 specific (ACC-049, Alomone Labs).

Techniques: Negative Control

Journal: Molecular Neurodegeneration

Article Title: TRPM8 and Na v 1.8 sodium channels are required for transthyretin-induced calcium influx in growth cones of small-diameter TrkA-positive sensory neurons

doi: 10.1186/1750-1326-6-19

Figure Lengend Snippet: TrkA- and TRPM8 expression and morphology of DRG neurons in culture . (A&B) Small diameter DRG neurons in culture express the TrkA receptor and TRPM8 channels. Immunofluoresence images of 12 hr DRG cultures stained with (A) an anti-TrkA (green), (B) an anti-TRPM8 (green) antibodies, and DAPI (blue). Analysis of somatic diameters displaying immunoreactivity (closed circles) for (C) TrkA and (D) TRPM8 show a significant bias of TrkA and TRPM8 immunoreactivity (closed circles) in smaller neurons (<15 μm) compared with TrkA and TRPM8 negative neurons (open circles). (n = 326 and 223 cells counted respectively, scale bar is 20 μm). (E) TRPM8 immunoreactivity in culture. DRG neurons displaying effusive growth cone morphologies (suitable of calcium imaging experiments) were TRPM8 positive (scale bar is 50 μm).

Article Snippet: The primary antibodies, a polyclonal rabbit anti-TRPM8 (extracellular) (1:1000, Alomone Labs, Israel) or polyclonal rabbit anti-TrkA (1:1000, Abcam, Cambridge, UK) were added to coverslips and incubated for 4 hr at 22°C.

Techniques: Expressing, Staining, Imaging

Journal: Molecular Neurodegeneration

Article Title: TRPM8 and Na v 1.8 sodium channels are required for transthyretin-induced calcium influx in growth cones of small-diameter TrkA-positive sensory neurons

doi: 10.1186/1750-1326-6-19

Figure Lengend Snippet: siRNA silencing of TRPM8 expression . Figure shows the level of TRPM8 immunoreactivity as determined by immunocytochemistry and by western blotting after treatment with siRNA oligonucleotides. (A) TRPM8 staining of a DRG growth cone treated with a non-specific control siRNA or with a specific TRPM8 siRNA oligonucleotide (#57381). The dotted line describes outline of the growth cone in the TRPM8 siRNA culture. (B) Quantitative analysis of TRPM8 immunocytochemistry shows that there was a significant decrease in TRPM8 expression in the presence of the TRPM8 siRNA (#57381, n = 73 growth cones) compared with the non-specific control siRNA (n = 56 growth cones). (C,D) Western blot and quantitation of TRPM8 immunoreactivity after treatment with 3 different TRPM8 siRNA oligonucleotides and a non-specific control siRNA. (C) Extracts from siRNA-treated DRG cultures were applied to SDS gels and probed with an anti-TRPM8 antibody. (D) Quantitative analysis of blots revealed significant knockdown of TRPM8 protein expression compared to cultures treated with a non-specific control siRNA. TRPM8 siRNA levels were normalised to GAPDH expression. Figure shows 60-70% knockdown achieved with 3 separate TRPM8 oligonucleotides. Values are means of 9 replicate immunoblot lanes over 3 separate experiments. Significant differences from control are depicted as: ** p < 0.005 as determined by a Mann-Whitney U-test. Error bars show means ± SEM

Article Snippet: The primary antibodies, a polyclonal rabbit anti-TRPM8 (extracellular) (1:1000, Alomone Labs, Israel) or polyclonal rabbit anti-TrkA (1:1000, Abcam, Cambridge, UK) were added to coverslips and incubated for 4 hr at 22°C.

Techniques: Expressing, Immunocytochemistry, Western Blot, Staining, Quantitation Assay, MANN-WHITNEY

Journal: Molecular Neurodegeneration

Article Title: TRPM8 and Na v 1.8 sodium channels are required for transthyretin-induced calcium influx in growth cones of small-diameter TrkA-positive sensory neurons

doi: 10.1186/1750-1326-6-19

Figure Lengend Snippet: Effect of TRPM8 knockdown on TTR-induced calcium influx . TRPM8 channels are necessary for L55P-induced calcium influx. Representative images of DRG neurons loaded with (A) non-specific control siRNA elicited a greater L55P-induced calcium influx (ΔF/F 0 ) than growth cones loaded with (B) specific TRPM8 siRNA (#57381). (C) Pooled results show calcium influx (ΔF/F 0 ) in response to L55P for control siRNA (closed circles, n = 12) and a specific TRPM8 siRNA (open circles, n = 16) from 3 separate experiments. (D) Using 3 different TRPM8 siRNA oligonucleotides results in a similar reduction in L55P-mediated calcium entry. Significant differences from control are depicted as: *p < 0.05; **p < 0.005; Mann-Whitney U-test. Error bars indicate mean ± SEM. Scale bar is 5 μm.

Article Snippet: The primary antibodies, a polyclonal rabbit anti-TRPM8 (extracellular) (1:1000, Alomone Labs, Israel) or polyclonal rabbit anti-TrkA (1:1000, Abcam, Cambridge, UK) were added to coverslips and incubated for 4 hr at 22°C.

Techniques: MANN-WHITNEY

Journal: PLoS ONE

Article Title: Antibodies to the Extracellular Pore Loop of TRPM8 Act as Antagonists of Channel Activation

doi: 10.1371/journal.pone.0107151

Figure Lengend Snippet: Alignment of the third extracellular loop sequences of the human, rat and mouse TRPM8 channel and the human TRPA1 and TRPV1 channels. The red line indicates the epitope sequence that ACC-049 was generated against.

Article Snippet: ACC-049, a rabbit polyclonal TRPM8 antibody generated against an epitope in the third extracellular loop near the pore region of human TRPM8 was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Sequencing, Generated

Journal: PLoS ONE

Article Title: Antibodies to the Extracellular Pore Loop of TRPM8 Act as Antagonists of Channel Activation

doi: 10.1371/journal.pone.0107151

Figure Lengend Snippet: Specificity of ACC-049 (2.5 µM) for blocking human TRPM8 activation induced by the specific natural agonist cold (A) or synthetic agonist icilin (D). No effect of ACC-049 on noxious cold induced human TRPA1 (B) or heat induced TRPV1 activation (C). Small molecule antagonists AMG9090 and AMG6541 are the positive control for TRPA1 (B) or TRPV1 (C) blockage, respectively. Note the near complete blockade of TRPM8 activation by ACC-049 at 2.5 µM, similar to that by the positive small molecule antagonist control M8-B (A). Neither control IgG, nor peptide-absorbed ACC-049, or peptide alone blocked activation of any of the channels tested (A–D). Values are means of triplicate measures in a single experiment and expressed as percent of control (POC). Agonist induced 45 Ca 2+ uptake in the absence of antibodies (no Ab) was considered as 100 percent and wells with small molecule antagonists plus 45 Ca 2+ were set as zero percent.

Article Snippet: ACC-049, a rabbit polyclonal TRPM8 antibody generated against an epitope in the third extracellular loop near the pore region of human TRPM8 was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Blocking Assay, Activation Assay, Positive Control

Journal: PLoS ONE

Article Title: Antibodies to the Extracellular Pore Loop of TRPM8 Act as Antagonists of Channel Activation

doi: 10.1371/journal.pone.0107151

Figure Lengend Snippet: Concentration dependent antagonism of cold activation (10°C) of the human (A), rat (B), or mouse (C) TRPM8 channels by ACC-049, control IgG and M8-B measured by 45 calcium uptake. Note the right shifted concentration response of ACC-049 on human TRPM8 (A) compared to rat (B) or mouse (C) TRPM8, while the small molecule antagonist positive control M8-B exhibited comparable responses on TRPM8 channels of all species tested (A–C). Values are means of triplicate measures in a single experiment and expressed as percent of control (POC). Cold induced 45 Ca 2+ uptake was considered as 100 percent and wells with M8-B at 1 µM plus 45 Ca 2+ were set as zero percent.

Article Snippet: ACC-049, a rabbit polyclonal TRPM8 antibody generated against an epitope in the third extracellular loop near the pore region of human TRPM8 was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Concentration Assay, Activation Assay, Positive Control

Journal: PLoS ONE

Article Title: Antibodies to the Extracellular Pore Loop of TRPM8 Act as Antagonists of Channel Activation

doi: 10.1371/journal.pone.0107151

Figure Lengend Snippet: IC 50 values (nM) of cold, icilin, and menthol induced human, rat, or mouse TRPM8 channel activation by ACC-049.

Article Snippet: ACC-049, a rabbit polyclonal TRPM8 antibody generated against an epitope in the third extracellular loop near the pore region of human TRPM8 was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Activation Assay

Journal: PLoS ONE

Article Title: Antibodies to the Extracellular Pore Loop of TRPM8 Act as Antagonists of Channel Activation

doi: 10.1371/journal.pone.0107151

Figure Lengend Snippet: Concentration dependent antagonism of icilin induced activation of the human (A), rat (B) or mouse (C) TRPM8 channels by ACC-049, control IgG and M8-B measured by 45 calcium uptake. Note the right shifted concentration response of ACC-049 on human TRPM8 (A) compared to rat (B) or mouse (C) TRPM8, while the small molecule antagonist positive control M8-B exhibited comparable responses on TRPM8 channels of all species tested (A–C). Values are means of triplicate measures in a single experiment and expressed as percent of control (POC). Icilin induced 45 Ca 2+ uptake was considered as 100 percent and wells with only assay buffer plus 45 Ca 2+ were set as zero percent.

Article Snippet: ACC-049, a rabbit polyclonal TRPM8 antibody generated against an epitope in the third extracellular loop near the pore region of human TRPM8 was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Concentration Assay, Activation Assay, Positive Control

Journal: PLoS ONE

Article Title: Antibodies to the Extracellular Pore Loop of TRPM8 Act as Antagonists of Channel Activation

doi: 10.1371/journal.pone.0107151

Figure Lengend Snippet: Concentration dependent antagonism of menthol induced activation of the human (A), rat (B) or mouse (C) TRPM8 channels by ACC-049, control IgG and M8-B measured by 45 calcium uptake. Human TRPM8 channel activation was blocked by ACC-049 in a concentration dependent manner (A), but there was no antagonistic effect of ACC-049 on either rat (B) or mouse (C) TRPM8 channels activated by menthol. The small molecule antagonist positive control M8-B exhibited comparable responses on TRPM8 channels of all species tested (A–C). Values are means of triplicate measures and expressed as percent of control (POC). Menthol induced 45 Ca 2+ uptake was considered as 100 percent and wells with only assay buffer plus 45 Ca 2+ were set as zero percent.

Article Snippet: ACC-049, a rabbit polyclonal TRPM8 antibody generated against an epitope in the third extracellular loop near the pore region of human TRPM8 was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Concentration Assay, Activation Assay, Positive Control

Journal: PLoS ONE

Article Title: Antibodies to the Extracellular Pore Loop of TRPM8 Act as Antagonists of Channel Activation

doi: 10.1371/journal.pone.0107151

Figure Lengend Snippet: Antagonism of icilin induced activation of human TRPM8 recombinantly expressed by CHO cells (A) and rat DRG neurons (B) by additional poly- and monoclonal antibodies generated against the third extracellular pore loop. a . Alomone ACC-049. b . MyBiosource MBS609041. c . Creative Diagnostics CABT37242RH. d. Thermo Scientific OST00133W. e . Antibodies Online ABIN351226. f . Lifespan Biosciences LS-B6668. g . Enzo Lifesciences BML-SA664. h . M8-B. i . 1 µM icilin. j . 1 µM icilin + peptide (SDVD GTTYDFAHC). k . Buffer. A. Note the complete block of TRPM8 channel activation by ACC-049 ( a ), MyBiosource ( b ) and Enzo Lifesciences ( g ) antibodies at the single concentration tested. Small molecule positive control M8-B also completely blocked TRPM8 channel activation ( h ). B. Five out of seven antibodies tested ( a, b, e, f, g ) block icilin activation of rat DRG neurons by 70–80%, two antibodies ( c, d ) are ineffective. Values are means of triplicate measures in a single experiment and expressed as percent of control (POC). 45 Ca 2+ uptake of CHO-TRPM8 cells activated with 1 µM icilin and antigen peptide ( j ) was considered as 100 percent and wells with only assay buffer plus 45 Ca 2+ were set as zero percent.

Article Snippet: ACC-049, a rabbit polyclonal TRPM8 antibody generated against an epitope in the third extracellular loop near the pore region of human TRPM8 was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Activation Assay, Generated, Blocking Assay, Concentration Assay, Positive Control

Journal: Current biology : CB

Article Title: Growth at Cold Temperature Increases the Number of Motor Neurons to Optimize Locomotor Function

doi: 10.1016/j.cub.2019.04.072

Figure Lengend Snippet: (A) RT-PCR for trpm8 (236 bp) from cDNA extracted from stage 24 whole embryo or dissected spinal cord. +/− RT, in the presence or absence of the reverse transcriptase, respectively, during conversion of isolated mRNA into cDNA. (B) In situ hybridization for trpm8 in stage 24 embryos showing specific labeling in brain and spinal cord. (C) Western blot assays from egg, stage 24 wild-type whole embryo or from stage 40 whole larva, control morpholino (MO) or TRPM8-translation-blockingMO 1 (TRPM8-tbMO1) lysates. Predicted TRPM8 molecular weight [MW]: 132 kDa. Shown are representative examples of one of 3 independent experiments. β-tubulin was used as loading control. (D) Immunostained transverse section of stage 25 spinal cord (outlined). D, dorsal; V, ventral; scale bar, 20 μm; arrows indicate TRPM8 clusters in ventral neuron domains. NCAM labeling was used as counterstaining. (E) Stage 24 ventral spinal cord from wild-type embryos was Ca 2+ imaged at 1 Hz for 90 s. Either 100 μM (−)-menthol or vehicle (0.05% DMSO)was added after 35 s of imaging and recording continued for another 60 s. Images show a menthol-responsive ventral neuron before (left, control) and after (right) addition of (−)-menthol. Colored scale shows fluorescence intensity in arbitrary units. Traces show the changes in fluorescence for the indicated cell (arrow) in both trials. (F) Stage 24 ventral spinal cord from wild-type embryos was Ca 2+ imaged in 30-min intervals at cold (14.5°C) and warm (26.5°C) temperatures in the absence (vehicle, 0.1% DMSO) or presence of 10 μM AMTB, TRPM8 inhibitor. Scatterplots show changes in Ca 2+ spike frequency when switching temperatures in individual spinal neurons and geometric mean (black lines) from N = 3 ventral spinal cords per condition (n of neurons analyzed: DMSO, 62; AMTB, 71). Teal circles represent neurons with higher spike frequency at 14.5°C, magenta circles represent neurons with higher spike frequency at 26.5°C, and black circles represent neurons with no change in spike frequency across temperatures; ****p < 0.0001, comparison within treatments Wilcoxon matched-pairs signed rank, two-tailed test. (G) RT-PCR from cDNA collected from stage 46 larvae previously injected with 2.5 pmol standard control morpholino (Control-MO) or TRPM8-splicing-blocking morpholino (TRPM8-sbMO) shows that trpm8 mature transcript (349 bp) is not detected in TRPM8-sbMO animals. odc : ornithine decarboxylase (101 bp) as positive control. (H) TRPM8-sbMO or Control-MO containing spinal cord from stage 24 embryos were Ca 2+ -imaged for 30 min at cold temperature (14.5°C). Graph shows individual (scatterplots) and geometric mean (black lines) Ca 2+ spike frequency from N = 3 ventral spinal cords pergroup (n of neurons analyzed: Control-MO, 81; TRPM8-sbMO, 64), ****p < 0.001, Kolmogorov-Smirnov, two-tailed test. See also and .

Article Snippet: Immunoblotting was performed overnight with anti-TRPM8 rabbit polyclonal affinity-purified antibody (1:500-1:700 in 5% milk) raised against the peptide SDVDGTTYDFAHC, corresponding to the amino acid residues 917-929 in the 3 rd extracellular loop of human TRPM8 (Alomone Labs), which is conserved (11 out of 13 residues) in Xenopus laevis TRPM8.

Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, In Situ Hybridization, Labeling, Western Blot, Molecular Weight, Imaging, Fluorescence, Two Tailed Test, Injection, Blocking Assay, Positive Control

Journal: Current biology : CB

Article Title: Growth at Cold Temperature Increases the Number of Motor Neurons to Optimize Locomotor Function

doi: 10.1016/j.cub.2019.04.072

Figure Lengend Snippet: (A) Images show cross-sections of immunostained spinal cord (outlined) for HB9 from Control-MO- or TRPM8-sbMO-containing stage 40 larvae grown in cold temperature (16°C). Scale bar, 20 μm. (B) Graph shows number of HB9-immunopositive cells per 100 μm of spinal cord; mean ± SEM from at least 220 μm-length spinal cord per larva, n = 5 larvae per condition, **p < 0.005, two-tailed t test. (C-G) TRPM8-sbMO or Control-MO stage 37/38 (C) and 40 (D–G) larvae grown in cold temperature (16°C) were subjected to 20 trials each of gentle touch at cold (14.5°C) or warm (22.5°C) temperature. Responses were video recorded at 30 Hz, n = 6 larvae per condition. (C) Average percentage of incidence of each response. (D) Time course of changes in speed during initial phase (first 200 ms) of the swim response. (E) Swim duration. (F) Maximum speed during initial phase of the swim response. (G) Maximum tangential acceleration during initial phase of the swim response. In (D)–(G), data are mean ± SEM. In (C) and (E)–(G), the letters on top of datasets indicate significant (different letters) or not significant (same letters) differences, p < 0.05, 2-way ANOVA, Tukey post hoc test. See also .

Article Snippet: Immunoblotting was performed overnight with anti-TRPM8 rabbit polyclonal affinity-purified antibody (1:500-1:700 in 5% milk) raised against the peptide SDVDGTTYDFAHC, corresponding to the amino acid residues 917-929 in the 3 rd extracellular loop of human TRPM8 (Alomone Labs), which is conserved (11 out of 13 residues) in Xenopus laevis TRPM8.

Techniques: Two Tailed Test

Journal: Current biology : CB

Article Title: Growth at Cold Temperature Increases the Number of Motor Neurons to Optimize Locomotor Function

doi: 10.1016/j.cub.2019.04.072

Figure Lengend Snippet: TRPM8 activation at cold temperature results in an increase in Ca 2+ spike frequency in developing spinal cord neurons. This enhanced spike frequency increases HB9-dependent motor neuron differentiation and survival. The low-temperature mediated increase in motor neuron number facilitates faster escape swimming at cold temperature compared to animals grown in warm temperature, for efficiently evading predators and increasing rate of survivability.

Article Snippet: Immunoblotting was performed overnight with anti-TRPM8 rabbit polyclonal affinity-purified antibody (1:500-1:700 in 5% milk) raised against the peptide SDVDGTTYDFAHC, corresponding to the amino acid residues 917-929 in the 3 rd extracellular loop of human TRPM8 (Alomone Labs), which is conserved (11 out of 13 residues) in Xenopus laevis TRPM8.

Techniques: Activation Assay

Journal: Current biology : CB

Article Title: Growth at Cold Temperature Increases the Number of Motor Neurons to Optimize Locomotor Function

doi: 10.1016/j.cub.2019.04.072

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Immunoblotting was performed overnight with anti-TRPM8 rabbit polyclonal affinity-purified antibody (1:500-1:700 in 5% milk) raised against the peptide SDVDGTTYDFAHC, corresponding to the amino acid residues 917-929 in the 3 rd extracellular loop of human TRPM8 (Alomone Labs), which is conserved (11 out of 13 residues) in Xenopus laevis TRPM8.

Techniques: Recombinant, Western Blot, In Situ, Luciferase, Reporter Assay, Isolation, Sequencing, Software

Journal: Current biology : CB

Article Title: Growth at Cold Temperature Increases the Number of Motor Neurons to Optimize Locomotor Function

doi: 10.1016/j.cub.2019.04.072

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-TRPM8 (extracellular) , Alomone Labs , Cat# ACC-049; RRID: AB_2040254.

Techniques: Recombinant, Western Blot, In Situ, Luciferase, Reverse Transcription, cDNA Synthesis, Isolation, Sequencing, Control, Knockdown, Software