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Choline Acetyltransferase/ChAT Antibody (334008)

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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.
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 ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB
Clone
334008
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
LYOPH

Order Details

View Available Formulations
Catalog# & Formulation Size Price
Carrier Free
MAB3447-SP
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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
Innovator's Reward
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Applications/Dilutions

Dilutions
  • Western Blot 1 ug/mL
Publications
Read Publications using
MAB3447 in the following applications:

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.

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Publications for Choline Acetyltransferase/ChAT Antibody (MAB3447)(4)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: ELISA Capture.


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Secondary Antibodies

 

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Blogs on Choline Acetyltransferase/ChAT.

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Choline Acetyltransferase (ChAT) – a useful Immunohistochemical marker for morphological studies of neurons
Choline Acetyltransferase (ChAT) is the enzyme that is responsible for biosynthesis of the neurotransmitter acetylcholine. The majority of acetylcholine is synthesized locally at nerve terminals where ChAT catalyzes the transfer of an acetyl group ...  Read full blog post.
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Bioinformatics

Gene Symbol CHAT
Uniprot