trpa1  (Novus Biologicals)

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Calcium imaging and immunocytochemistry showing TRPV1 expression in transfected HEK‐293 cells. (A) Fura2 calcium imaging in HEK‐293 cells transiently expressing rTRPV1‐EYFP fusion protein. Representative traces of calcium transients evoked by capsaicin in EYFP + cells. Green traces correspond to EYFP + cells and gray traces to EYFP − cells. Pie chart represents the percentage of EYFP + cells that responded or failded to respond to capsaicin. Data obtained from 88 cells from 3 coverslips. (B–F left) Confocal images of TRPV1‐EYFP transiently expressed in HEK‐293 cells. EYFP (green), TRPV1 antibody (magenta) and Hoechst (HO, blue). Merge images correspond to the overlap between TRPV1 and EYFP on the left and Hoechst and brightfield (BF) on the right. Antibody dilution of the representative images is indicated in the lower right corner. Scale bar: 20 μm. (B–F right) Box plots display the specificity ratio (SR) for each tested dilution of the corresponding antibody. Each dot represents a single EYFP + cell. Boxes indicate the interquartile range (25th to 75th percentiles) and whiskers extend to the 5th and 95th percentiles. The horizontal line inside each box marks the median and the black dot indicates the mean. (*** p < 0.001 Mann–Whitney test). (G) Bar histogram graph summarizes the SR median difference between the tested antibody dilution and their controls lacking primary antibody (secondary antibody only; rabbit, goat or guinea pig) as calculated with the Hodges‐Lehman estimator. Error bars represent the 95% confidence interval. For each antibody and dilution, a minimum of 100 cells were analyzed across 4 fields from 2 independent transfections. $ Santa Cruz V1 antibody was generated against a human TRPA1 peptide.

Article Snippet: Alomone , TRPA1 , Rabbit , Alomone Labs , Human TRPA1 747–760. PC , 69.2% , ACC‐037 RRID:AB_2040232.

Techniques: Imaging, Immunocytochemistry, Expressing, Transfection, MANN-WHITNEY, Generated

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Calcium imaging and immunocytochemistry of TRPA1 expression in transfected HEK‐ 293 cells. (A) Representative traces of Fura2 calcium transients evoked by 50 μM AITC. Green traces correspond to tGFP + cells and gray traces to tGFP − cells. The pie chart represents the proportion of tGFP + cells that responded or failed to respond to AITC. Data obtained from 115 cells from 3 coverslips. (B–H left) Confocal images of TRPA1‐tGFP transiently expressed in HEK‐ 293 cells incubated with the indicated antibody. tGFP (green), TRPA1 (magenta) and Hoechst staining (blue). Merge images correspond to the overlap between TRPA1 and tGFP (lower‐left panel) or Hoechst and brightfield (HO, BF, lower‐right). Antibody dilution of the representative images is indicated in the lower right‐hand corner. Scale bar: 20 μm. (B–H right) box plots display the specificity ratio (SR) for each tested dilution of the corresponding antibody. Each dot represents a single tGFP + cell. Boxes indicate the interquartile range (25th to 75th percentiles) and whiskers extend to the 5th and 95th percentiles. The horizontal line inside each box marks the median and the black dot indicates the mean. (** p < 0.01, *** p < 0.001 Mann–Whitney test). (I) Bar histogram graph summarizes the SR median difference between the tested antibody dilution and their controls lacking primary antibody (secondary antibody only; rabbit or mouse) as calculated with the Hodges‐Lehman estimator. Error bars represent the 95% confidence interval. For each antibody and dilution, a minimum of 319 cells were analyzed across 4 fields from 2 independent transfections.

Article Snippet: Alomone , TRPA1 , Rabbit , Alomone Labs , Human TRPA1 747–760. PC , 69.2% , ACC‐037 RRID:AB_2040232.

Techniques: Imaging, Immunocytochemistry, Expressing, Transfection, Incubation, Staining, MANN-WHITNEY

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Western blot analysis for TRPV1 and TRPA1 antibodies specificity. (A) Immunoblots for TRPV1 using Abcam (1:1000), Millipore V1 (1:1000), Santa Cruz V1 (1:100), Neuromics GT (1:200), and Neuromics GP (1:1000) antibodies. (−) lanes: Untransfected HEK‐293 cells, (+) lanes: HEK‐293 cells transfected with rTRPV1‐EYFP (B) Immunoblots for TRPA1 using Aviva (1:300), Novus (1:500), Millipore A1 (1:500), Alomone (1:200), Proteintech (1:500), Sigma WH (1:500), and Santa Cruz A1 (1:100). (−) lanes: Untransfected HEK‐293 cells, (+) lanes: HEK‐293 cells transfected with hTRPA1‐tGFFP. For each antibody, top row: Immunoblot with each TRPV1/TRPA1 antibody. Middle row: EYFP/tGFP immunoblotting of the same membrane. Bottom image: GAPDH loading control. Each blot was performed at least three times to rule out technical artifacts in cases where no anti‐TRPV1/TRPA1 signal was detected. For every repetition, the identical lysate sample was probed with all antibodies. Full uncropped blots are available in the Figure . $ Santa Cruz V1 antibody was generated against a human TRPA1 peptide.

Article Snippet: Alomone , TRPA1 , Rabbit , Alomone Labs , Human TRPA1 747–760. PC , 69.2% , ACC‐037 RRID:AB_2040232.

Techniques: Western Blot, Transfection, Membrane, Control, Generated

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Calcium imaging and Immunochemistry of endogenous TRPA1 in cultured DRG neurons from TRPA1‐Cre‐ChR2‐EYFP mice. (A) Representative traces of calcium transients evoked by AITC and capsaicin in DRG neurons. Traces corresponding to cells responding to 50 μM AITC are displayed in yellow and non‐responding cells are displayed in gray. (B) The pie chart represents the proportion of neurons responding or failing to respond to AITC. Data obtained from 424 cells from 5 coverslips. (C–F) ICC or (G–J) IHC confocal images of DRG sensory neurons incubated with the indicated antibody. TRPA1 antibody (magenta) and βIIITubulin (cyan), merge images display TRPA1 antibody signal + βIII‐Tubulin. Dilution of the corresponding antibody shown in each example is indicated in the lower right corner. Scale bar: 50 μm. Representative images were chosen from 4 pictures of 2 different animals.

Article Snippet: Alomone , TRPA1 , Rabbit , Alomone Labs , Human TRPA1 747–760. PC , 69.2% , ACC‐037 RRID:AB_2040232.

Techniques: Imaging, Cell Culture, Incubation

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Calcium imaging and immunocytochemistry showing TRPV1 expression in transfected HEK‐293 cells. (A) Fura2 calcium imaging in HEK‐293 cells transiently expressing rTRPV1‐EYFP fusion protein. Representative traces of calcium transients evoked by capsaicin in EYFP + cells. Green traces correspond to EYFP + cells and gray traces to EYFP − cells. Pie chart represents the percentage of EYFP + cells that responded or failded to respond to capsaicin. Data obtained from 88 cells from 3 coverslips. (B–F left) Confocal images of TRPV1‐EYFP transiently expressed in HEK‐293 cells. EYFP (green), TRPV1 antibody (magenta) and Hoechst (HO, blue). Merge images correspond to the overlap between TRPV1 and EYFP on the left and Hoechst and brightfield (BF) on the right. Antibody dilution of the representative images is indicated in the lower right corner. Scale bar: 20 μm. (B–F right) Box plots display the specificity ratio (SR) for each tested dilution of the corresponding antibody. Each dot represents a single EYFP + cell. Boxes indicate the interquartile range (25th to 75th percentiles) and whiskers extend to the 5th and 95th percentiles. The horizontal line inside each box marks the median and the black dot indicates the mean. (*** p < 0.001 Mann–Whitney test). (G) Bar histogram graph summarizes the SR median difference between the tested antibody dilution and their controls lacking primary antibody (secondary antibody only; rabbit, goat or guinea pig) as calculated with the Hodges‐Lehman estimator. Error bars represent the 95% confidence interval. For each antibody and dilution, a minimum of 100 cells were analyzed across 4 fields from 2 independent transfections. $ Santa Cruz V1 antibody was generated against a human TRPA1 peptide.

Article Snippet: Novus , TRPA1 , Rabbit , Novus Biologicals (Biotechne) , Human TRPA1 N‐terminus (1–100) PC , 77% , NB110‐40763 RRID:AB_715124.

Techniques: Imaging, Immunocytochemistry, Expressing, Transfection, MANN-WHITNEY, Generated

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Calcium imaging and immunocytochemistry of TRPA1 expression in transfected HEK‐ 293 cells. (A) Representative traces of Fura2 calcium transients evoked by 50 μM AITC. Green traces correspond to tGFP + cells and gray traces to tGFP − cells. The pie chart represents the proportion of tGFP + cells that responded or failed to respond to AITC. Data obtained from 115 cells from 3 coverslips. (B–H left) Confocal images of TRPA1‐tGFP transiently expressed in HEK‐ 293 cells incubated with the indicated antibody. tGFP (green), TRPA1 (magenta) and Hoechst staining (blue). Merge images correspond to the overlap between TRPA1 and tGFP (lower‐left panel) or Hoechst and brightfield (HO, BF, lower‐right). Antibody dilution of the representative images is indicated in the lower right‐hand corner. Scale bar: 20 μm. (B–H right) box plots display the specificity ratio (SR) for each tested dilution of the corresponding antibody. Each dot represents a single tGFP + cell. Boxes indicate the interquartile range (25th to 75th percentiles) and whiskers extend to the 5th and 95th percentiles. The horizontal line inside each box marks the median and the black dot indicates the mean. (** p < 0.01, *** p < 0.001 Mann–Whitney test). (I) Bar histogram graph summarizes the SR median difference between the tested antibody dilution and their controls lacking primary antibody (secondary antibody only; rabbit or mouse) as calculated with the Hodges‐Lehman estimator. Error bars represent the 95% confidence interval. For each antibody and dilution, a minimum of 319 cells were analyzed across 4 fields from 2 independent transfections.

Article Snippet: Novus , TRPA1 , Rabbit , Novus Biologicals (Biotechne) , Human TRPA1 N‐terminus (1–100) PC , 77% , NB110‐40763 RRID:AB_715124.

Techniques: Imaging, Immunocytochemistry, Expressing, Transfection, Incubation, Staining, MANN-WHITNEY

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Western blot analysis for TRPV1 and TRPA1 antibodies specificity. (A) Immunoblots for TRPV1 using Abcam (1:1000), Millipore V1 (1:1000), Santa Cruz V1 (1:100), Neuromics GT (1:200), and Neuromics GP (1:1000) antibodies. (−) lanes: Untransfected HEK‐293 cells, (+) lanes: HEK‐293 cells transfected with rTRPV1‐EYFP (B) Immunoblots for TRPA1 using Aviva (1:300), Novus (1:500), Millipore A1 (1:500), Alomone (1:200), Proteintech (1:500), Sigma WH (1:500), and Santa Cruz A1 (1:100). (−) lanes: Untransfected HEK‐293 cells, (+) lanes: HEK‐293 cells transfected with hTRPA1‐tGFFP. For each antibody, top row: Immunoblot with each TRPV1/TRPA1 antibody. Middle row: EYFP/tGFP immunoblotting of the same membrane. Bottom image: GAPDH loading control. Each blot was performed at least three times to rule out technical artifacts in cases where no anti‐TRPV1/TRPA1 signal was detected. For every repetition, the identical lysate sample was probed with all antibodies. Full uncropped blots are available in the Figure . $ Santa Cruz V1 antibody was generated against a human TRPA1 peptide.

Article Snippet: Novus , TRPA1 , Rabbit , Novus Biologicals (Biotechne) , Human TRPA1 N‐terminus (1–100) PC , 77% , NB110‐40763 RRID:AB_715124.

Techniques: Western Blot, Transfection, Membrane, Control, Generated

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Calcium imaging and Immunochemistry of endogenous TRPA1 in cultured DRG neurons from TRPA1‐Cre‐ChR2‐EYFP mice. (A) Representative traces of calcium transients evoked by AITC and capsaicin in DRG neurons. Traces corresponding to cells responding to 50 μM AITC are displayed in yellow and non‐responding cells are displayed in gray. (B) The pie chart represents the proportion of neurons responding or failing to respond to AITC. Data obtained from 424 cells from 5 coverslips. (C–F) ICC or (G–J) IHC confocal images of DRG sensory neurons incubated with the indicated antibody. TRPA1 antibody (magenta) and βIIITubulin (cyan), merge images display TRPA1 antibody signal + βIII‐Tubulin. Dilution of the corresponding antibody shown in each example is indicated in the lower right corner. Scale bar: 50 μm. Representative images were chosen from 4 pictures of 2 different animals.

Article Snippet: Novus , TRPA1 , Rabbit , Novus Biologicals (Biotechne) , Human TRPA1 N‐terminus (1–100) PC , 77% , NB110‐40763 RRID:AB_715124.

Techniques: Imaging, Cell Culture, Incubation

Journal: bioRxiv

Article Title: Integrated transcriptomics and proteomics define the TRP channel hierarchy in mouse cortex

doi: 10.64898/2026.04.07.716663

Figure Lengend Snippet: ( A ) Bulk cortex TaqMan qPCR showing mean Ct values for Trpa1 and Trpv1 in adult mouse cortex. Both transcripts are detected only at high Ct values. Red dashed line indicates Gapdh Ct. ( B ) TaqMan qPCR of FACS-sorted cortical cell populations showing mean Ct values by fraction: neurons (blue), astrocytes (orange), and NeuN⁺/ACSA2⁺ double-positive cells (grey). Dashed lines indicate fraction-specific Gapdh Ct. “‡” indicate no detectable amplification reactions. Trpv1 is neuron-only at high Ct; Trpa1 is rare/borderline. ( C, D ) Subcellular fractionation and immunoblotting for TRPA1 and TRPV1, respectively. Cortex lysates (n = 3) were separated into supernatant (S) and membrane pellet (P) fractions; Ladder (L). Faint immunoreactive bands near the expected molecular weight (∼130–140 and ∼95 kDa, respectively) are preferentially enriched in pellet fractions. GAPDH (∼37 kDa) was used as a loading control. Kidney, DRG, and testes processed in parallel serve as positive control tissues. Note: Lysates and controls were processed in the same experiment and run on the same gels/blots. ( E, F ) Immunoprecipitation (IP) from cortex (C) and DRG (D) lysates followed by SDS–PAGE/Western blotting and LC–MS/MS. As shown in Supplementary Fig. 12f, no TRPA1 peptides were detected, and only extremely low-intensity TRPV1 protein-group signals were observed without replicate consistency.

Article Snippet: In the first approach, sections were incubated overnight at 4 °C with primary antibodies against TRPA1 (Alomone Labs, ACC-037) or TRPV1 (Alomone Labs, ACC-030), diluted 1:200 in blocking buffer.

Techniques: Amplification, Fractionation, Western Blot, Membrane, Molecular Weight, Control, Positive Control, Immunoprecipitation, SDS Page, Liquid Chromatography with Mass Spectroscopy