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, FC |
| Clonality: | Monoclonal |
| Clone number: | JA67-11 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| 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: | SH-SY5Y cell lysate, Neuro-2a cell lysate, Mouse brain tissue lysate, Mouse cerebellum tissue lysate, Rat brain tissue lysate, Rat cerebellum tissue lysate, HeLa. |
| Subcellular location: | Cytosol, nucleus, cytoplasm, neuron projection, presynapse. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:5,000 1:50-1:200 1:1,000 Use at an assay dependent concentration. |
| 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
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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: SH-SY5Y cell lysate (20 µg/Lane) Lane 2: Neuro-2a cell lysate (negative) (20 µg/Lane) Lane 3: Mouse brain tissue lysate (40 µg/Lane) Lane 4: Mouse cerebellum tissue lysate (40 µg/Lane) Lane 5: Rat brain tissue lysate (40 µg/Lane) Lane 6: Rat cerebellum tissue lysate (40 µg/Lane) Predicted band size: 83 kDa Observed band size: 50/75 kDa Exposure time: 5 seconds; ECL: K1801; 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 primary antibody dilution (K1803) at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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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. |
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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). |
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Fig4:
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). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.