PRODUCT CODE: ET1704-16

Recombinant Choline Acetyltransferase Monoclonal Antibody (ET1704-16)

  • Recombinant

Applications

  • WB

  • IP

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

ICC staining of Choline Acetyltransferase in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Choline Acetyltransferase in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Choline Acetyltransferase in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Choline Acetyltransferase in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Choline Acetyltransferase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-16, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded rat stomach tissue using anti-Choline Acetyltransferase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-16, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of Choline Acetyltransferase was done on N2A cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1704-16, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ICC staining of Choline Acetyltransferase in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-16, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Applications

  • WB

  • IP

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Choline Acetyltransferase Monoclonal Antibody (ET1704-16)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Hela, N2A, SH-SY5Y, mouse brain tissue, rat stomach tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JA67-11

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

82 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • 1:10-1:50

TARGET

UNIPROT #

PROTEIN NAME

Choline Acetyltransferase

SYNONYMS

Acetyl CoA choline O acetyltransferase antibody; Acetyl CoA:choline O acetyltransferase antibody; ChAT antibody; CHOACTase antibody; Choline acetylase antibody; choline acetyltransferase antibody; Choline O acetyltransferase antibody; Choline O-acetyltransferase antibody; CLAT_HUMAN antibody; CMS1A antibody; CMS1A2 antibody; EC 2.3.1.6 antibody; OTTHUMP00000019583 antibody; OTTHUMP00000019584 antibody

SEQUENCE SIMILARITIES

Belongs to the carnitine/choline acetyltransferase family.

SUBCELLULAR LOCATION

Cytosol, nucleus, cytoplasm, neuron projection, presynapse

FUNCTION

Choline acetyltransferase (also designated choactase, choline O-acetyltransferase) synthesizes acetylcholine in cholinergic neurons. Multiple choactase mRNAs with different 5'-noncoding regions are expressed as R-, N1, N2-, S- and M-types. N1-, N2- and R-type mRNAs produce a single short enzyme, while M-type mRNA produces both long and short enzymes. The long enzyme is targeted to the nuclei of cells, whereas the short protein is found in cytoplasm. A novel NFkB binding site is located within the nerve growth factor-responsive enhancer element that is recognized by the NFkB protein p49, but not p65 or p50. Decreased choactase expression and increased NFkB activity are associated with aging and Alzheimer's disease, indicating that p49 is a negative regulator of choactase expression and suggesting a possible mechanism for aging-associated declines in cholinergic function. Phosphorylation of choactase has been shown to enhance choactase catalytic activity. Specifically, Serine 440 is found to be the phosphorylation site in a recombinant human short choactase by protein kinase C and is involved in regulation of the enzyme catalytic activity and binding to subcellular membranes.