MAB305 Anti-Choline Acetyltransferase Antibody, clone 1E6

MAB305
100 µL  
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, Mk, R IHC M Ascites Monoclonal Antibody
      Description
      Catalogue NumberMAB305
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAnti-Choline Acetyltransferase Antibody, clone 1E6
      Alternate Names
      • ChAT
      • Choline Acetylase
      • CHOACTase
      References
      Product Information
      FormatAscites
      Control
      • Brain tissue
      PresentationAscites fluid containing no preservatives.
      Quality LevelMQ100
      Applications
      ApplicationDetect Choline Acetyltransferase using this Anti-Choline Acetyltransferase Antibody, clone 1E6 validated for use in IH.
      Key Applications
      • Immunohistochemistry
      Application NotesImmunohistochemistry: 1:100-1:250. See immunohistochmistry procedure below.

      Optimal working dilutions must be determined by the end user.

      IMMUNOHISTOCHEMISTRY PROCEDURE (PAP TECHNIQUE) FOR MAB305, MONOCLONAL ANTIBODY TO CHOLINE ACETYLTRANSFERASE

      I) Perfusion & Sectioning Procedure

      1. Perfuse through the heart with a fixative solution containing 4% paraformaldehyde in 0.12 M phosphate buffer (pH 7.3) for light microscopy (LM), and additionally, 0.1% gluteraldehyde and .002% CaCl2 for electron microscopy (EM).

      2. Remove brain and postfix 2-18 hours at 4°C in 4% paraformaldehyde in 0.12 M phosphate buffer.

      3. After brain is blocked for sectioning, wash in several changes of buffer for 2-3 hours.

      4. Specimens for EM are sectioned on a Vibratome (50 μm) and rinsed in buffer, those for LM should be cryoprotected in 30% sucrose in buffer.

      5. After freezing with dry ice, 30-40 μm thick sections of LM specimens are cut on a cryostat.

      6. Sections are rinsed, and then stored in phosphate buffer containing 0.1% sodium azide.

      II) Staining Procedure

      Tissue is processed as freely-floating sections in continuously agitated solutions. All incubations are performed at room temperature unless otherwise stated.

      1.a. For localizing ChAT-positive somata and dendrites:

      Sections are washed in 0.1 M Tris-buffered saline (TBS; containing 1.4% NaCl, pH 7.3) only. No detergent or enzyme pretreatment is used.

      b. For localizing ChAT-positive terminal-like structures:

      Incubate sections in TBS (pH 8.1) for 5 minutes at 37°C. Transfer sections to TBS (pH 8.1) containing pronase (1.2 μg/mL) for 1 1/2-2 minutes at 37°C, followed by several ice cold buffer washes for a total of 5 minutes. The concentration of pronase and incubation time of the digestion should be evaluated for each region examined.

      c. For localizing ChAT immunoreactivity and subsequently counterstaining the sections:

      Incubation in TBS containing 0.1%-0.8% Triton X-100 for 15 minutes may increase the tissue penetration of the immunoreagents, but it also raises the background staining.

      2. Incubate sections in normal goat serum (3-5%) for one hour. The working solutions of all antisera should also contain similarly diluted normal goat serum.

      3. Incubate in anti-ChAT monoclonal antibody solution (Suggested working dilution 1:250, final working dilution must be determined by end user) for 2 hours at room temperature and then for an additional 6-18 hours at 4°C.

      4. Incubate with second antibody (i.e. Goat anti-Mouse IgG, Cat. No.: AP124, dilution 1:50-100) for 1-2 hours.

      5. Incubate with diluted PAP complex (i.e. Mouse PAP, Cat No.: PAP14, conc. 25-50 μg/mL) for one hour.

      6. After rinsing in buffer, the second antibody and PAP steps are repeated for 40 minutes to 1 hour each in order to amplify staining intensity, particularly of small ChAT-containing structures.

      7. React for 15 minutes with 0.06% 3,3'-diaminobenzidine×4 HCl (DAB; diluted in phosphate buffered saline, pH 7.3) and 0.006% H2O2.

      8. Specimens for routine LM are postfixed for 1 minutes in 0.005% OsO4 (osmium tetraoxide), and then mounted, dehydrated and coverslipped. Selected regions blocked for EM are postfixed in 2% OsO4 for 1 hour, en bloc stained with uranyl acetate, and flat-embedded in Epon-Araldite resin.
      Biological Information
      ImmunogenCholine acetyltransferase purified from rat brain.
      Clone1E6
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificityRecognizes cholinergic neurons in the brain and spinal cord (CNS).
      IsotypeIgG1
      Species Reactivity
      • Human
      • Monkey
      • Rat
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryCholinergic systems are implicated in numerous neurologic functions. Alteration in some cholinergic neurons may account for the disturbances of Alzheimer disease. The protein encoded by this gene synthesizes the neurotransmitter acetylcholine. Alternative splice variants have been found that contain alternative 5' untranslated exons. Three of the four described splice variants encode identical 69 kDa proteins while one variant encodes both the 69 kDa and a larger 82 kDa protein.
      Gene Symbol
      • CHAT
      • ChAT
      • CMS1A2
      • CHOACTase
      • CMS1A
      • EC 2.3.1.6
      Purification MethodUnpurified
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P28329 # Catalyzes the reversible synthesis of acetylcholine (ACh) from acetyl CoA and choline at cholinergic synapses.
      SIZE: 748 amino acids; 82568 Da
      DISEASE: SwissProt: P28329 # Defects in CHAT are the cause of familial infantile myasthenia gravis 2 (FIMG2) [MIM:254210, 254200]; also known as CMS-EA. FIMG2 patients have myasthenic symptoms since birth or early infancy, negative tests for anti-AChR antibodies, and abrupt episodic crises with increased weakness, bulbar paralysis, and apnea precipitated by undue exertion, fever, or excitement. Inheritance is autosomal recessive.
      SIMILARITY: SwissProt: P28329 ## Belongs to the carnitine/choline acetyltransferase family.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsMaintain for 1 year at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Anti-Choline Acetyltransferase Antibody, clone 1E6 SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-Choline Acetyltransferase Antibody, clone 1E6 Certificates of Analysis

      TitleLot Number
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) MONOCLONAL ANTIBODY - 21200502120050
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) MONOCLONAL ANTIBODY - 21400092140009
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) MONOCLONAL ANTIBODY - 23707362370736
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) MONOCLONAL ANTIBODY - 24556492455649
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) -26104592610459
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) -26215022621502
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) -26766582676658
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) -27244552724455
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) -27754922775492
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) -28026512802651

      References

      Reference overviewApplicationSpeciesPub Med ID
      Glucocorticoid receptors in the locus coeruleus mediate sleep disorders caused by repeated corticosterone treatment.
      Wang, ZJ; Zhang, XQ; Cui, XY; Cui, SY; Yu, B; Sheng, ZF; Li, SJ; Cao, Q; Huang, YL; Xu, YP; Zhang, YH
      Scientific reports  5  9442  2015

      Show Abstract
      Immunohistochemistry25801728 25801728
      microRNA-125 distinguishes developmentally generated and adult-born olfactory bulb interneurons.
      Akerblom, M; Petri, R; Sachdeva, R; Klussendorf, T; Mattsson, B; Gentner, B; Jakobsson, J
      Development (Cambridge, England)  141  1580-8  2014

      Show Abstract
      Immunohistochemistry24598163 24598163
      Wnts enhance neurotrophin-induced neuronal differentiation in adult bone-marrow-derived mesenchymal stem cells via canonical and noncanonical signaling pathways.
      Tsai, HL; Deng, WP; Lai, WF; Chiu, WT; Yang, CB; Tsai, YH; Hwang, SM; Renshaw, PF
      PloS one  9  e104937  2014

      Show Abstract
      ImmunofluorescenceHuman25170755 25170755
      Characterization of cognitive deficits in rats overexpressing human alpha-synuclein in the ventral tegmental area and medial septum using recombinant adeno-associated viral vectors.
      Hall, H; Jewett, M; Landeck, N; Nilsson, N; Schagerlöf, U; Leanza, G; Kirik, D
      PloS one  8  e64844  2013

      Show Abstract
      Immunohistochemistry23705016 23705016
      Infrared optical imaging of matrix metalloproteinases (MMPs) up regulation following ischemia reperfusion is ameliorated by hypothermia.
      Barber, PA; Rushforth, D; Agrawal, S; Tuor, UI
      BMC neuroscience  13  76  2012

      Show Abstract
      22742423 22742423
      Salubrinal, an endoplasmic reticulum stress blocker, modulates sleep homeostasis and activation of sleep- and wake-regulatory neurons.
      Methippara, M, et al.
      Neuroscience, 209: 108-18 (2012)  2012

      Show Abstract
      22387272 22387272
      Increased numbers of motor activity peaks during light cycle are associated with reductions in adrenergic alpha(2)-receptor levels in a transgenic Huntington's disease rat model.
      Bode FJ, Stephan M, Wiehager S, Nguyen HP, Bjorkqvist M, von Horsten S, Bauer A, Petersen A
      Behavioural brain research  205  175-182  2009

      Show Abstract
      19573560 19573560
      Brain injury does not alter the intrinsic differentiation potential of adult neuroblasts.
      Liu, F; You, Y; Li, X; Ma, T; Nie, Y; Wei, B; Li, T; Lin, H; Yang, Z
      The Journal of neuroscience : the official journal of the Society for Neuroscience  29  5075-87  2009

      Show Abstract
      ImmunohistochemistryRat19386903 19386903
      Activated cholinergic signaling provides a target in squamous cell lung carcinoma.
      Song, P; Sekhon, HS; Fu, XW; Maier, M; Jia, Y; Duan, J; Proskosil, BJ; Gravett, C; Lindstrom, J; Mark, GP; Saha, S; Spindel, ER
      Cancer research  68  4693-700  2008

      Show Abstract Full Text Article
      18559515 18559515
      Neonatal hypoxic/ischemic brain injury induces production of calretinin-expressing interneurons in the striatum.
      Zhengang Yang,Yan You,Steven W Levison
      The Journal of comparative neurology  511  2008

      Show Abstract Full Text Article
      18720478 18720478

      Data Sheet

      Title
      MOUSE ANTI-CHOLINE ACETYLTRANSFERASE (ChAT) MONOCLONAL ANTIBODY

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