trpm8 polyclonal antibody  (Proteintech)

Journal: International Journal of Molecular Sciences

Article Title: Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression

doi: 10.3390/ijms232416164

Figure Lengend Snippet: Immunocytochemistry of mTRPM8 in transfected HEK-293 cells. ( A – F left) Confocal images of mTRPM8-EYFP transiently expressed in HEK-293 cells. EYFP (green), TRPM8 antibody (red) and Hoechst staining (blue). Merge images correspond to the overlap of the three fluorescent signals plus the bright field image. Scale bar: 30 µm. ( A – F right) Box plots represent the specificity ratio (SR) for each dilution of the respective antibody. Each dot corresponds to an individual EYFP+ cell. Box contains the 25th to 75th percentiles. Whiskers mark the 5th and 95th percentiles. The line inside the box denotes the median and the black dot represents the mean. (*** p < 0.001, Mann–Whitney test). ( G ) Bar histogram summarizing the SR mean ± SEM of each TRPM8 antibody and the controls without primary antibody (αRabbit or αMouse secondary antibodies alone). Dashed line indicates SR = 1. (*** p < 0.001, Kruskal–Wallis followed by Dunn’s post hoc test vs. αRabbit or αMouse). For each antibody and dilution, n > 270 cells; 4 fields from 2 independent transfections.

Article Snippet: TRPM8 , Origene1 , OriGene , TA307827 , Rabbit , Primary , Human TRPM8 extracellular domain. Monoclonal , 1:200 (ICC, ICH) 1:500 (WB, ICC, ICH).

Techniques: Immunocytochemistry, Transfection, Staining, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression

doi: 10.3390/ijms232416164

Figure Lengend Snippet: Western blot analysis for TRPM8 antibodies specificity. ( A – F ) TRPM8 immunoblots using ( A ) Alomone ( B ) ECM1, ( C ) Origene1, ( D ) ECM2, ( E ) ECM3 or ( F ) Origene2 antibodies. (−) lanes: untransfected HEK-293 cells, (+) lanes: HEK-293 cells transfected with mTRPM8-EYFP. Left: Immunoblot with each TRPM8 antibody. Right-top: EYFP immunoblotting on the same membrane. Right-bottom: GAPDH loading control. Black arrowheads indicate mTRPM8-EYFP bands revealed with antiTRPM8 antibody. Green arrowheads indicate mTRPM8-EYFP bands revealed with anti-GFP antibody. Red arrowheads indicate GAPDH bands. All blots were repeated at least 3 times to exclude a technical artefact when no anti-TRPM8 signal was observed. For each replicate, the same lysate was used for all antibodies.

Article Snippet: TRPM8 , Origene1 , OriGene , TA307827 , Rabbit , Primary , Human TRPM8 extracellular domain. Monoclonal , 1:200 (ICC, ICH) 1:500 (WB, ICC, ICH).

Techniques: Western Blot, Transfection, Membrane, Control

Journal: International Journal of Molecular Sciences

Article Title: Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression

doi: 10.3390/ijms232416164

Figure Lengend Snippet: Calcium imaging in DRG-cultured cells from Trpm8 BAC -EYFP + mice. ( A ) Representative traces of calcium transients evoked by mild cold (∼18–20 °C), WS-12 (10 µM) and KCl (30 mM) in EYFP+ and EYFP− neurons. Traces corresponding to individual cells are shown in grey. Colored traces represent averages of the respective individual traces. ( B ) Proportion of EYFP+ and EYFP− cells responding to cold and WS-12. ( C ) Representative immunocytochemistry performed after a calcium recording. Upper- and bottom-left: confocal images of the immunofluorescence. Green: EYFP. Red: TRPM8 (ECM1 antibody). Grey: βIII-Tubulin. Bottom-right: transmitted light and EYFP fluorescence from the same live cells before fixation. n = 432 cells from 2 mice. Scale bar: 50 µm.

Article Snippet: TRPM8 , Origene1 , OriGene , TA307827 , Rabbit , Primary , Human TRPM8 extracellular domain. Monoclonal , 1:200 (ICC, ICH) 1:500 (WB, ICC, ICH).

Techniques: Imaging, Cell Culture, Immunocytochemistry, Immunofluorescence, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression

doi: 10.3390/ijms232416164

Figure Lengend Snippet: Immunocytochemistry of endogenous TRPM8 in cultured DRG cells from the Trpm8 BAC -EYFP + mouse. ( A – F left) Confocal images of TRPM8+ sensory neurons. EYFP (green), TRPM8 antibody (red) and βIII-Tubulin (grey). Scale bar: 50 µm. ( A – F right) Box plots represent specificity ratio (SR) for each dilution of the respective antibody. Each dot corresponds to an individual EYFP+ cell. Box contains the 25th to 75th percentiles. Whiskers mark the 5th and 95th percentiles. The line inside the box denotes the median and the black dot represents the mean (* p < 0.05, ** p < 0.01, Mann–Whitney test). ( G ) Bar histogram summarizing the SR mean ± SEM of each TRPM8 antibody and the controls without primary antibody (αRabbit or αMouse secondary antibodies alone). Dashed line indicates SR = 1 (* p < 0.05, ** p < 0.01, Kruskal–Wallis followed by Dunn’s post hoc test vs. αRabbit or αMouse). n = at least 20 cells; 4 pictures from 2 mice for each antibody and dilution.

Article Snippet: TRPM8 , Origene1 , OriGene , TA307827 , Rabbit , Primary , Human TRPM8 extracellular domain. Monoclonal , 1:200 (ICC, ICH) 1:500 (WB, ICC, ICH).

Techniques: Immunocytochemistry, Cell Culture, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression

doi: 10.3390/ijms232416164

Figure Lengend Snippet: Immunohistochemistry of endogenous TRPM8 in DRG slices from the Trpm8 BAC -EYFP + mouse. ( A – F left) Confocal images of TRPM8+ sensory neurons. EYFP (green), TRPM8 antibody (red) and βIII-Tubulin (grey). Scale bar: 50 µm. ( A – F right) Box plots represent specificity ratio (SR) for each dilution of the respective antibody. Each dot corresponds to an individual EYFP+ cell. Box contains the 25th to 75th percentiles. Whiskers mark the 5th and 95th percentiles. The line inside the box denotes the median and the black dot represents the mean. (Differences among dilutions were not significant, Mann–Whitney test). ( G ) Bar histogram summarizing the SR mean ± SEM of each TRPM8 antibody and the controls without primary antibody (αRabbit or αMouse secondary antibodies alone). Dashed line indicates SR = 1. (* p < 0.05, ** p < 0.01, Kruskal–Wallis followed by Dunn´s post hoc test vs. αRabbit or αMouse). n = at least 18 cells; 4 images from 2 mice for each antibody and dilution.

Article Snippet: TRPM8 , Origene1 , OriGene , TA307827 , Rabbit , Primary , Human TRPM8 extracellular domain. Monoclonal , 1:200 (ICC, ICH) 1:500 (WB, ICC, ICH).

Techniques: Immunohistochemistry, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression

doi: 10.3390/ijms232416164

Figure Lengend Snippet: Immunofluorescence of endogenous TRPM8 in DRG cells and slices from the TRPM8 KO mouse. ( A , C ) Immunocytochemistry. ( B , D ) Immunohistochemistry. ( A – D left) Confocal images of TRPM8 KO ( Trpm8 EGFPf ; B6;129S1(FVB)- Trpm8 tm1Apat /J) sensory neurons. EGFP (green), TRPM8 antibody (red) and βIII-Tubulin (grey). Scale bar: 50 µm. ( A – D right) Box plots represent specificity ratio (SR) for each dilution of the respective antibody. Each dot corresponds to an individual EGFP+ cell. Box contains the 25th to 75th percentiles. Whiskers mark the 5th and 95th percentiles. The line inside the box denotes the median and the black dot represents the mean (** p < 0.01, Mann–Whitney test). ( E,F ) Bar histograms summarizing the SR mean ± SEM of each TRPM8 antibody in the KO and the reporter ( Trpm8 BAC -EYFP + , named M8-EYFP in the figure) mouse in ICC ( E ) and IHC ( F ). Dashed line indicates mean SR for the control without primary antibody (** p < 0.01, Mann–Whitney test). n = at least 22 cells; 4 pictures from 2 mice for each antibody and dilution.

Article Snippet: TRPM8 , Origene1 , OriGene , TA307827 , Rabbit , Primary , Human TRPM8 extracellular domain. Monoclonal , 1:200 (ICC, ICH) 1:500 (WB, ICC, ICH).

Techniques: Immunofluorescence, Immunocytochemistry, Immunohistochemistry, MANN-WHITNEY, Control

Journal: International Journal of Molecular Sciences

Article Title: Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression

doi: 10.3390/ijms232416164

Figure Lengend Snippet: Antibody performance with the different techniques used in this study. − poor, + regular, ++ good, +++ excellent.

Article Snippet: TRPM8 , Origene1 , OriGene , TA307827 , Rabbit , Primary , Human TRPM8 extracellular domain. Monoclonal , 1:200 (ICC, ICH) 1:500 (WB, ICC, ICH).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression

doi: 10.3390/ijms232416164

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: TRPM8 , Origene1 , OriGene , TA307827 , Rabbit , Primary , Human TRPM8 extracellular domain. Monoclonal , 1:200 (ICC, ICH) 1:500 (WB, ICC, ICH).

Techniques: Isolation