TRPA1 Antibody

Immunocytochemistry/Immunofluorescence: TRPA1 Antibody [NB110-40763] - Analysis of TRPA1 in HEK 293 cells (Flp In Trex system) cells.

Immunohistochemistry: TRPA1 Antibody [NB110-40763] - Staining TRPA1 in mouse intestine.

Flow Cytometry: TRPA1 Antibody [NB110-40763] - An intracellular stain was performed on A549 cells with NB110-40763C (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 650.

TRPA1 Antibody [NB110-40763] - An intracellular stain was performed on A549 cells with NB110-40763G (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 488

• Reactivity: Hu, Mu, Rt, GP
• Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

TRPA1 Antibody Summary

• Immunogen: A synthetic peptide made to a region within the N-terminus (residues 1-100) of the human TRPA1 protein. [Swiss-Prot# O75762]
• Localization: Membrane
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TRPA1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 2 ug/ml
  • ELISA
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:200
  • Immunohistochemistry 0.5-1.0 ug/ml
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:100-1:250
• Application Notes: This TRPA1 antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry paraffin embedded sections and Western blot analysis, where a band is seen at ~110 kDa. Immunohistochemistry-Frozen was reported in scientific literature. Use in FLOW reported in scientific literature (PMID: 28990934). Use in ELISA reported in scientific literature (PMID: 29803505).

Reactivity Notes

Immunogen sequence has 69% homology to mouse. Rat reactivity reported in scientific literature (PMID: 27748654).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for TRPA1 Antibody

• ANKTM1ankyrin-like with transmembrane domains 1
• Ankyrin-like with transmembrane domains protein 1
• Transformation-sensitive protein p120
• transient receptor potential cation channel subfamily A member 1
• transient receptor potential cation channel, subfamily A, member 1

Background

TRPA1 (transient receptor potential cation channel subfamily A member 1; originally named as ANKTM1) is a member of transient receptor potential ion channels (TRPCs) which responds to stimuli such as cold temperatures, pungent compounds, and plays role in both nociceptor and hair cell transductions. TRPA1 is activated by variety of mechanical and chemicals stimuli including dietary irritants such as isothyocyanates (found in mustard oil, horse-radish, wasabi), allycin/garlic etc. and its activity is regulated by endogenous pro-inflammatory factors such as cyclopentane prostaglandins and byproducts of oxidative stress (e.g. 4-HNE, 4-oxononenal) which activate TRPA1 directly via covalent modification of its cysteine residues. Once TRPA1 is activated, increased intracellular Ca2+ induces the peripheral release of neuropeptides namely substance P and CGRP (calcitonin gene-related peptide), purines, and other transmitters from sensitized nerve endings, which results in neurogenic inflammation and hypersensitivity. TRPA1 is mainly expressed in small-diameter peptidergic nociceptors of the dorsal root, nodose and trigeminal ganglia, along with TRPV1, and has also been reported to be expressed in non-neuronal tissues (skeletal muscle, lung, small intestine, colon, pancreas etc) and is found dysregulated during process of carcinogenesis in certain malignancies.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for TRPA1 Antibody (NB110-40763)(30)

Publications using NB110-40763

• Brandolini L, Castelli V, Aramini A et al. DF2726A, a new IL-8 signalling inhibitor, is able to counteract chemotherapy-induced neuropathic pain Sci Rep Aug 13 2019 [PMID: 31409858] (WB, Rat)
• Ma S, Zhang Y, He K et al. Knockout of TRPA1 exacerbates angiotensin II-induced kidney injury Am. J. Physiol. Renal Physiol. Jul 24 2019 [PMID: 31339777]
• Miao, F;Wang, R;Cui, G;Li, X;Wang, T;Li, X; Engagement of MicroRNA-155 in Exaggerated Oxidative Stress Signal and TRPA1 in the Dorsal Horn of the Spinal Cord and Neuropathic Pain During Chemotherapeutic Oxaliplatin Neurotox Res Apr 23 2019 [PMID: 31016687] (WB, Rat)
• Kameda, T;Zvick, J;Vuk, M;Sadowska, A;Tam, WK;Leung, VY;Bolcskei, K;Helyes, Z;Applegate, LA;Hausmann, ON;Klasen, J;Krupkova, O;Wuertz-Kozak, K; Expression and Activity of TRPA1 and TRPV1 in the Intervertebral Disc: Association with Inflammation and Matrix Remodeling Int J Mol Sci Apr 10 2019 [PMID: 30974795] (ICC/IF)
• Conklin DJ, Guo Y, Jagatheesan G et al. Transient Receptor Potential Ankyrin-1 (TRPA1) Channel Contributes to Myocardial Ischemia-Reperfusion Injury Am. J. Physiol. Heart Circ. Physiol. Feb 8 2019 [PMID: 30735434] (IHC-P, Mouse)
• Ma W, Chen X, Cerne R et al. Catechol estrogens stimulate insulin secretion in pancreatic b-cells via activation of TRPA1 channel J. Biol. Chem. Dec 26 2018 [PMID: 30587572] (WB, Human)
• Peixoto-Neves D, Soni H, Adebiyi A. CGRPergic Nerve TRPA1 Channels Contribute to Epigallocatechin Gallate-Induced Neurogenic Vasodilation ACS Chem Neurosci Dec 4 2018 [PMID: 30513192] (ICC/IF, Mouse)
• Wang Z, Xu Y, Wang M et al. TRPA1 inhibition ameliorates pressure overload-induced cardiac hypertrophy and fibrosis in mice bioRxiv Oct 17 2018 [PMID: 30297144] (Human)
• Takahashi N, Chen HY, Harris IS et al. Cancer Cells Co-opt the Neuronal Redox-Sensing Channel TRPA1 to Promote Oxidative-Stress Tolerance Cancer Cell May 15 2018 [PMID: 29805077] (IHC)
• Lowin T, Bleck J, Schneider M, Pongratz G. Selective killing of proinflammatory synovial fibroblasts via activation of transient receptor potential ankyrin (TRPA1) Biochem. Pharmacol. May 24 2018 [PMID: 29803505] (WB, ICC/IF, ELISA, Human)
• De Logu F, Nassini R, Materazzi S et al. Schwann cell TRPA1 mediates neuroinflammation that sustains macrophage-dependent neuropathic pain in mice. Nat Commun 2017 Dec 01 [PMID: 29192190] (Mouse)
• Matsuda K, Okamoto N, Kondo M et al. Mast cell hyperactivity underpins the development of oxygen-induced retinopathy. J. Clin. Invest. 2017 Nov 01 [PMID: 28990934] (Flow, Mouse)
• Nummenmaa E, Hamalainen M, Moilanen LJ et al. TRPA1 expression is downregulated by dexamethasone and aurothiomalate in human chondrocytes: TRPA1 as a novel factor and drug target in arthritis RMD Open 2017 Sep 15 [PMID: 28912961] (WB, Human)
• Chen JL, Ping YH, Tseng MJ et al. Notch1-promoted TRPA1 expression in erythroleukemic cells suppresses erythroid but enhances megakaryocyte differentiation. Sci Rep. 2017 Feb 21 [PMID: 28220825] (WB, Human)
• Hidaka T, Ogawa E, Kobayashi EH et al. The aryl hydrocarbon receptor AhR links atopic dermatitis and air pollution via induction of the neurotrophic factor artemin. Nat. Immunol. 2016 Nov 21 [PMID: 27869817] (Mouse)
  
  Details:
  This citation used the DyLight 650 version of this antibody.
• Lu Y, Piplani H, McAllister SL et al. Transient Receptor Potential Ankyrin 1 Activation within the Cardiac Myocyte Limits Ischemia-reperfusion Injury in Rodents Anesthesiology. 2016 Dec 01 [PMID: 27748654] (WB, Rat)
• Nummenmaa Elina, Hamalainen Mari, Moilanen Lauri J et al. Transient receptor potential ankyrin 1 (TRPA1) is functionally expressed in primary human osteoarthritic chondrocytes. Arthritis Research & Therapy 2016 [PMID: 27515912] (WB, Human)
• Lee KI, Lin HC, Lee HT et al. Loss of Transient Receptor Potential Ankyrin 1 Channel Deregulates Emotion, Learning and Memory, Cognition, and Social Behavior in Mice. Mol. Neurobiol. May 19 2016 [PMID: 27194300] (WB, Mouse)
• Lee KI, Lee HT, Lin HC et al. Role of transient receptor potential ankyrin 1 channels in Alzheimer's disease. J Neuroinflammation. Apr 28 2016 [PMID: 27121378] (WB, IHC, Mouse, Human)
• Takaya J, Mio K, Shiraishi T et al. A Potent and Site-Selective Agonist of TRPA1. J. Am. Chem. Soc. 2015 Dec 23 [PMID: 26630251] (Human)
• Billeter AT, Galbraith N, Walker S et al. TRPA1 mediates the effects of hypothermia on the monocyte inflammatory response. Surgery 2015 Jun 5 [PMID: 26054320]
• Weng HJ, Patel KN, Jeske NA et al. Tmem100 Is a Regulator of TRPA1-TRPV1 Complex and Contributes to Persistent Pain Neuron. 2015 Jan 28 [PMID: 25640077] (WB, Mouse)
• Terada Y, Fujimura M, Nishimura S et al. Roles of Cav 3.2 and TRPA1 channels targeted by hydrogen sulfide in pancreatic nociceptive processing in mice with or without acute pancreatitis. J. Neurosci. Res. 2014 Sep 30 [PMID: 25267397] (WB, Mouse)
  
  Details:
  TRPA1 antibody used for WB in experiments involving mouse model of acute pancreatitis. Figure 5 shows TRPA1 expression in dorsal root ganglia of mice with TRPA1 knockdown achieved via repeated intrathecal administration of the mixture of two or three distinct oligodeoxynucleotides (ODNs) - AS-ODNs-TRPA1 (AS) or SC-ODNs-TRPA1 (SC). Figure 6 shows the expression levels of TRPA1 in the DRG of mice with cerulein / Cer-induced pancreatitis.
• Fusi C, Materazzi S, Minocci D et al. Transient Receptor Potential Vanilloid 4 (TRPV4) Is Downregulated in Keratinocytes in Human Non-Melanoma Skin Cancer. J. Invest. Dermatol. 2014 Mar 18 [PMID: 24643128] (IHC-P, Human)
• Oh MH, Oh SY, Lu J et al. TRPA1-Dependent Pruritus in IL-13-Induced Chronic Atopic Dermatitis. J Immunol. 2013 Dec 1 [PMID: 24140646] (IHC, ICC/IF, Mouse, Human)
• Okubo K, Matsumura M, Kawaishi Y et al. Hydrogen sulfide-induced mechanical hyperalgesia and allodynia require activation of both Cav3 2 and TRPA1 channels in mice. Br J Pharmacol 2012 Jul [PMID: 22300342]
  
  Details:
  Using the HRP conjugated version of NB110-40763, catalog number NB110-40763H.
• Mukhopadhyay I, Gomes P, Aranake S, Shetty M, Karnik P, Damle M, Kuruganti S, Thorat S, Khairatkar-Joshi N. Expression of functional TRPA1 receptor on human lung fibroblast and epithelial cells. J Recept Signal Transduct Res;31(5):350-8. 2011 Oct. [PMID: 21848366] (WB, Human)
• Poole DP, Pelayo JC, Cattaruzza F et al. Transient receptor potential ankyrin 1 is expressed by inhibitory motoneurons of the mouse intestine Gastroenterology 2011 Aug [PMID: 21689654] (IHC, ICC/IF, Mouse)
• Cattaruzza F, Spreadbury I, Miranda-Morales M et al. Transient receptor potential ankyrin-1 has a major role in mediating visceral pain in mice. Am J Physiol Gastrointest Liver Physiol 2010 Jan [PMID: 19875705] (IHC, ICC/IF, Mouse)
• Yu S, Gao G, Peterson BZ et al. TRPA1 in mast cell activation-induced long-lasting mechanical hypersensitivity of vagal afferent C-fibers in guinea pig esophagus. Am J Physiol Gastrointest Liver Physiol;297(1):G34-42. 2009 [PMID: 19423751] (ICC/IF, WB, IHC-Fr, Guinea Pig)

Review for TRPA1 Antibody (NB110-40763) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: TinaMarie Lieu IF Human 08/21/2012

Summary

• Application: Immunofluorescence
• Sample Tested: HEKFLPNTREX Cell line
• Species: Human
• Comments: TRP channel antibodies are difficult to stain with specificity. This TRPA1 antibody from Novus works pretty well.

Blocking

• Blocking Details: Block with 10% goat serum in 1% BSA & 0.1% Saponin for 2 hrs at RT

Primary Anitbody

• Dilution Ratio: Primary antibody of TRPA1 from Novus Biologicals – dilute in 1% BSA/0.1% Saponin (1:200 dilution)

Secondary Antibody

• Secondary Description: Secondary antibody diluted in 1%BSA/0.1% Saponin (1:1000 dilution) – Invitrogen Alexa Flour Goat anti-rabbit
• Secondary Concentration: 1:1000

Details

• Detection Notes: Leica with MetaFlour Flourescene Ratio Imaging, Pos. Control (DRG expressed TRPA1), Neg. Control (HEK nontransfected with TRPA1)
• Fixation Details: Fixed in cold methanol for 2 minutes (4C)
• Wash Description: PBS Wash 3X

Comments

• Comments: TRP channel antibodies are difficult to stain with specificity. This TRPA1 antibody from Novus works pretty well.

FAQs for TRPA1 Antibody (NB110-40763). (Showing 1 - 4 of 4 FAQs).

1. I read the first publication (Yu S et al. 2009) referenced for NB110-40763. This product reacts with human and mouse according to your datasheet, but in that publication it was used for western blot in guinea pig. Does this mean this antibody is suitable to use for guinea pig samples?
   • Yes, according to this paper this antibody has been successfully used on guinea pig samples. This is the only testing that has been done in guinea pig though as we have done no in-house testing. It should work just fine for you.
2. Has the specificity of NB110-40763 has been confirmed by mouse dorsal root ganglion (DRG) tissue in immunohistochemistry? Do you know what sample types have been tested?
   • We have a couple of publications with the base product catalog number NB110-40763 (which is just the unconjugated version of NB110-40763G), which have utilized our product with mouse colonic dorsal root ganglion. (PMID 19875705 and PMID 21689654). The image on our website was taken of a mouse intestine sample.
3. What are the other two bands in the image on the datasheet? UniProt finds a theoretical MW of 127.5kDa. You find a MW of 110kDa in WB application. There are two more bands at 60kDa and 17kDa. What are these? Unspecific staining or fragments or variants of the protein? I did not find any info on variants when looking into the UniProt data (http://www.uniprot.org/uniprot/O75762).
   • We generally use gradient gels in our lab and for proteins which are multi-pass membrane protein, it is not uncommon to see an approximate 10-15% change in expected vs observed molecular weight in WB assay. The band observed at approximately 110kD is highly specific and there is no doubt that the antibody is detecting the right band. No, we have not characterized the additional low molecular weight bands yet which are considerably mild bands when compared to the expected target band. During a brief literature research this morning, I came across some papers wherein other researchers have also observed additional lower molecular weight bands and some of those references are: J Biol Chem. 2010 May 14; 285(20): 15167-15177; J Biol Chem. 2006 October 27; 281(43): 32879-32890; J Clin Invest. 2007 July 2; 117(7): 1979-1987.
4. Human brain membrane fraction is mentioned to be used as a positive control for this antibody. Do you sell the positive control too?
   • The Western blot data that is shown on our product page for the TRPA1 antibody with catalogue number NB110-40763 was, as you have pointed out, generated from human brain membrane lysate. We do sell a human brain membrane fraction that is suitable for Western blotting, and you can see this here: TRPA1 antibody datasheet. This lysate is supplied as native protein, and we recommend adding 1 x sample buffer with 5% BME (or other reducing agent) prior to use.

Bioinformatics

• Gene Symbol: TRPA1
• Entrez:
  • 8989(Human)
• Uniprot:
  • O75762(Human)