Choline Acetyltransferase Recombinant Rabbit Monoclonal Antibody [JA67-11]
cat.: ET1704-16
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IP, IF-Cell, IF-Tissue, IHC-P, FC, IHC-Fr
Clonality: Monoclonal
Clone number: JA67-11
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 83 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Choline Acetyltransferase aa 699-748 / 748.
Positive control: Human brain tissue lysate, mouse brain tissue lysate, rat brain tissue lysate, Hela, N2A, SH-SY5Y, mouse brain tissue, rat stomach tissue, mouse hippocampus tissue, mouse cerebral cortex tissue.
Subcellular location: Cytosol, nucleus, cytoplasm, neuron projection, presynapse
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
  IHC-Fr
1:5,000
1:50-1:200
1:50-1:200
1:50-1:200
1:1,000
Use at an assay dependent concentration.
1:100
Uniprot #: SwissProt: P28329 Human | Q03059 Mouse | P32738 Rat
Alternative names: Acetyl CoA choline O acetyltransferase Acetyl CoA:choline O acetyltransferase ChAT CHOACTase Choline acetylase choline acetyltransferase Choline O acetyltransferase Choline O-acetyltransferase CLAT_HUMAN CMS1A CMS1A2 EC 2.3.1.6 OTTHUMP00000019583 OTTHUMP00000019584
Images
ET1704-16_1.jpg Fig1: Western blot analysis of Choline Acetyltransferase on different lysates with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16) at 1/5,000 dilution.

Lane 1: Human brain tissue lysate
Lane 2: Mouse brain tissue lysate
Lane 3: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 83 kDa
Observed band size: 60-83 kDa

Exposure time: 20 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-16) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1704-16_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling Choline Acetyltransferase with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1704-16_3.jpg Fig3: 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).
ET1704-16_4.jpg Fig4: 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).
ET1704-16_5.jpg Fig5: 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.
ET1704-16_6.jpg Fig6: 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.
ET1704-16_7.jpg Fig7: Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Choline Acetyltransferase with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16).

The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody ((ET1704-16, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
ET1704-16_8.jpg Fig8: Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Choline Acetyltransferase with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16).

The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody ((ET1704-16, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
ET1704-16_9.jpg Fig9: Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Choline Acetyltransferase with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16).

The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1704-16, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
ET1704-16_10.jpg Fig10: Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Choline Acetyltransferase with Rabbit anti-Choline Acetyltransferase antibody (ET1704-16).

The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1704-16, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
ET1704-16_11.jpg Fig11: Flow cytometric analysis of HeLa cells labeling Choline Acetyltransferase.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1704-16, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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