Choline Acetyltransferase/ChAT Antibody (334008)

Detection and characterization of extracellular ChAT protein in human plasma and CSF.(a) Dot-blot analysis of nine different pooled CSF samples using two different antibodies. (b) The relative amount of ChAT in plasma or CSF, compared to brain homogenates (BH) of AD or control. At 16-fold dilutions, the immunosignals are much stronger in plasma and CSF compared to BH. This demonstrates the relative abundance of the ChAT protein in extracellular fluids. (c) Western-blot characterization of molecular form of ChAT in plasma, CSF and BH. The major detected protein band (*) in the BHs corresponds to a ChAT protein with a Mw of ∼65 kDa. In addition, there are several detected heavier molecular forms of ChAT in the CSF. All samples were loaded on one gel. Note also that the amount of the ∼65 kDa ChAT was so high in plasma that it distorted the gel downward. (d–g) Combinatorial sandwich ELISA results for identification of extracellular ChAT using three different antibody pairs for recombinant human ChAT (rhChAT), and a pooled plasma sample (g) calibrated for ChAT protein concentration using the rhChAT and ELISA setup in (f). (h) Further characterization of the molecular forms of CSF ChAT in consecutive fractions of pooled AD CSF samples separated by sucrose-density gradient technique, and the subsequent quantification of ChAT by sandwich ELISA. The graph represents the average of nine different pooled CSF. This independent analysis provides an identical pattern of molecular forms detected by Western analysis. Due to lack of prior reports, we used analogous terminology, which is used for the counteracting cholinergic enzymes, AChE and BuChE, that is, Gn, where n denotes the number of globular subunits in each detected molecular form of ChAT in CSF. The molecular weights are calculated based on the known Mw of two internal standard proteins. In all dot-blot analyses, 2 µL of each sample (neat or diluted) was used. In the Western blot analysis, each lane was loaded with 15 µL of a mixture, containing 10 µL of sample (neat or diluted) and 5 µL of a 6x concentrated reducing Laemmli loading buffer. All ChAT protein quantifications were done with the ELISA antibody pair’s combination in (g). Anti-ChAT antibodies: RP = rabbit polyclonal antibody (Ab), BGP = biotin-labeled goat polyclonal Ab, MAB = mouse monoclonal Ab. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0065936), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB
• Clone: 334008
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Choline Acetyltransferase/ChAT Antibody (334008) Summary

• Immunogen: E. coli-derived recombinant human Choline Acetyltransferase/ChAT isoform 1
  Ala2-Pro630
  Accession # NP_066266
• Specificity: Detects human Choline Acetyltransferase/ChAT in direct ELISAs and Western blots. Recognizes both ChAT isoform 1 (ChAT-69) and isoform 2 (ChAT-82).
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: CHAT
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Choline Acetyltransferase/ChAT Antibody (334008)

• acetyl CoA:choline O-acetyltransferase
• CHAT
• CHOACTase
• Choline acetylase
• Choline Acetyltransferase
• choline O-acetyltransferase
• CMS1A
• CMS1A2
• EC 2.3.1
• EC 2.3.1.6

Background

ChAT catalyzes the production of the neurotransmitter acetylcholine (Ach), which is required for cholinogenic neuron communication. It serves as a marker for functional cholinogenic neurons in the central and peripheral nervous systems. At least six ChAT mRNAs encoding 69 kDa, 82 kDa and 74 kDa ChAT proteins have been identified. Compared to the 82 kDa form, 69 kDa ChAT lacks the N‑terminal 118 amino acid extension containing a nuclear localization signal. As a result the 69 kDa ChAT is primarily localized to the cytoplasm. Human 69 kDa ChAT shares 86% amino acid sequence identity with the mouse or rat ChAT.

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 Choline Acetyltransferase/ChAT Antibody (MAB3447)(5)

Publications using MAB3447

• Thakur, B;Tarazi, S;Doležalová, L;Behbahani, H;Darreh-Shori, T; Comparative Analysis of Cholinergic Machinery in Carcinomas: Discovery of Membrane-Tethered ChAT as Evidence for Surface-Based ACh Synthesis in Neuroblastoma Cells International journal of molecular sciences 2025-10-23 [PMID: 41226363] (Immunocytochemistry, Human)
• Zachary T. Olmsted, Cinzia Stigliano, Annalisa Scimemi, Tatiana Wolfe, Jose Cibelli, Philip J. Horner et al. Transplantable human motor networks as a neuron-directed strategy for spinal cord injury iScience 8/20/2021 [PMID: 34381965]
• Azadeh Karami, Maria Eriksdotter, Ahmadul Kadir, Ove Almkvist, Agneta Nordberg, Taher Darreh-Shori CSF Cholinergic Index, a New Biomeasure of Treatment Effect in Patients With Alzheimer’s Disease Frontiers in Molecular Neuroscience 10/11/2019 [PMID: 31680850]
• LI Sinclair, A Kumar, T Darreh-Sho, S Love Visual hallucinations in Alzheimer's disease do not seem to be associated with chronic hypoperfusion of to visual processing areas V2 and V3 but may be associated with reduced cholinergic input to these areas Alzheimers Res Ther, 2019-09-12;11(1):80. 2019-09-12 [PMID: 31511061] (ELISA Capture, Human)
• Swetha Vijayaraghavan, Azadeh Karami, Shahin Aeinehband, Homira Behbahani, Alf Grandien, Bo Nilsson et al. Regulated Extracellular Choline Acetyltransferase Activity— The Plausible Missing Link of the Distant Action of Acetylcholine in the Cholinergic Anti-Inflammatory Pathway PLoS ONE 6/19/2013 [PMID: 23840379]

Bioinformatics

• Gene Symbol: CHAT
• Uniprot:
  • NP_066266()