Anti trpa1

Manufactured by Novus Biologicals

Sourced in United States

Anti-TRPA1 is a lab equipment product used for the detection and quantification of the Transient Receptor Potential Ankyrin 1 (TRPA1) protein. TRPA1 is a member of the transient receptor potential (TRP) ion channel family and is involved in various physiological processes, including sensory perception and pain signaling. The Anti-TRPA1 product provides a tool for researchers to study the expression, localization, and function of TRPA1 in different biological systems.

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5 protocols using anti trpa1

Western Blot Protein Detection Protocol

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Western blot was done as previously described [19 (link)–21 (link)]. Primary antibodies used in the study were as follows: anti-TRPA1 (1:1000, Alomone), anti-TRPA1 (1:1000, Novus Biologicals, CO, USA), anti-TRPA1 (1:1000, LSBio, WA, USA), MAPK Family Antibody Sampler Kit (1:1000, Cell Signaling Technology), Phospho-MAPK Family Antibody Sampler Kit (1:1000, Cell Signaling Technology), anti-MKP-1 (1:1000, Thermo Fisher Scientific), PGC-1α (1:1000, Abcam), anti-β-actin (1:1000, Abcam), anti-β-tubulin (1:1000, Cell Signaling Technology). Secondary antibodies used were: HRP-conjugated goat anti-rabbit secondary antibody (1:3000, Dako), HRP-conjugated goat anti-mouse secondary antibody (1:3000, Dako).

Ding Q., Liu X., Qi Y., Yao X, & Tsang S.Y. (2023). TRPA1 promotes the maturation of embryonic stem cell-derived cardiomyocytes by regulating mitochondrial biogenesis and dynamics. Stem Cell Research & Therapy, 14, 158.

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Protein Expression Analysis of PDLCs

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The total protein of PDLCs and P-PDLCs or mouse gingiva was extracted, and 20 μg protein of each sample was separated and blotted onto a polyvinylidene fluoride membrane (Millipore, USA). Then, membranes were blocked and incubated with primary antibodies including anti-actin (Abcam, UK; ab3280, 1 : 10000), anti-cleaved Caspase-3 (Cell Signaling Technology, USA; #9664, 1 : 1000), anti- PPARγ2 (Abcam, UK; ab45036, 1 : 1000), anti-HSP70 (Zen, China; 200304, 1 : 1000), anti-TRPA1 (Novus, USA; NB100-91319, 1 : 1000), anti-SOD1 (Abcam, UK; ab51254, 1 : 1000), anti-SOD2 (Santa Cruz, USA; sc-133254, 1 : 1000), anti-PERK (Cell Signaling Technology, USA; #5683, 1 : 1000), anti-CHOP (Cell Signaling Technology, USA; #2895, 1 : 1000), anti-ATF-4 (Cell Signaling Technology, USA; #11815, 1 : 1000), anti-eIF2α (Cell Signaling Technology, USA; #5324, 1 : 1000), and anti-p-eIF2α (Cell Signaling Technology, USA; #3398, 1 : 1000) at 4°C overnight. The membranes were washed and incubated with secondary antibody (1 : 10,000) and were visualized by the Image Quant Tanon-5200Multi (Tanon, China). The relative intensity of the protein was quantitatively analyzed by the ratio of actin, and all assays were repeated three times.

Liu Q., Guo S., Huang Y., Wei X., Liu L., Huo F., Huang P., Wu Y, & Tian W. (2022). Inhibition of TRPA1 Ameliorates Periodontitis by Reducing Periodontal Ligament Cell Oxidative Stress and Apoptosis via PERK/eIF2α/ATF-4/CHOP Signal Pathway. Oxidative Medicine and Cellular Longevity, 2022, 4107915.

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Western Blotting of Cellular Proteins

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Whole-cell extracts were prepared and then resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis as described27 (link). Western blot analyses were performed with anti-Notch1 C terminal, anti-Ets-1, anti-DNMT3B (Santa Cruz), anti-cleaved Notch1, anti-ERK1/2, anti-phospho-ERK1/2 (Cell Signaling Technology), anti-TRPA1 (Novus Biologicals), anti-vimentin (Thermo Fisher Scientific), and anti-GAPDH (Genetex) antibodies.

Chen J.L., Ping Y.H., Tseng M.J., Chang Y.I., Lee H.C., Hsieh R.H, & Yeh T.S. (2017). Notch1-promoted TRPA1 expression in erythroleukemic cells suppresses erythroid but enhances megakaryocyte differentiation. Scientific Reports, 7, 42883.

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Intestinal Goblet and Enteroendocrine Cell Analysis

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Mouse colon was fixed in 10% neutral formalin and stored in 70% ethanol. Intestinal samples were then paraffin-embedded and cut into 10 um longitudinal sections, and the PAS staining was performed by Histoserv Inc. (Germantown, MD). Goblet cells were counted and calculated from 10 crypts in each slide.
For immunofluorescence staining, slides were deparaffinized by xylene and antigen retrieval was conducted for 20 min in a 95°-water bath in 10 mM sodium citrate, pH 6.0 followed by a 15 min incubation at room temperature. Slides were washed, blocked in 5% bovine serum albumin, and stained with the primary antibodies, anti-ChgA (1:100; Abcam), anti-5-HT (1:100; Abcam), anti-ST2 (1:100, Proteintech) and anti-TRPA1 (1:100, Novus) and secondary antibodies conjugated to Alexa fluor 488, 633 or 594 (Thermo Fisher). Slides were mounted in Fluoromount-G (Thermo Fisher), and 3–15 images were taken per slide at 20X or 40X magnification along transections of the intestinal crypts for each biological replicate (Zeiss). For 5-HT and ChgA staining, numbers of positively stained puncta were scored blindly, normalized to total area of intestinal mucosa using ImageJ software (NIH), and then averaged across biological replicates.

Chen Z., Luo J., Li J., Kim G., Stewart A., Urban JF J.r., Huang Y., Chen S., Wu L.G., Chesler A., Trinchieri G., Li W, & Wu C. (2020). Interleukin-33 promotes serotonin release from enterochromaffin cells for intestinal homeostasis. Immunity, 54(1), 151-163.e6.

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DRG Protein Expression Analysis

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DRG from cervical to lumbar levels were collected in 300 μL PBS and 2% SDS with protease inhibitor (Sigma P8340). The primary antibodies used were: 1:5000 anti-Tmem100, 1:1000 anti-TRPV1 (Santa Cruz, R130), and 1:1000 anti-TRPA1 (Novus, NB110-40763). Secondary antibodies for visualization included donkey ant-rabbit and anti-mouse HRP-conjugated antibodies (GE Biosciences). See supplementary data for details.

Weng H.J., Patel K.N., Jeske N.A., Bierbower S.M., Zou W., Tiwari V., Zheng Q., Tang Z., Mo G.C., Wang Y., Geng Y., Zhang J., Guan Y., Akopian A, & Dong X. (2015). Tmem100 is a regulator of TRPA1-TRPV1 complex and contributes to persistent pain. Neuron, 85(4), 833-846.

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