Anti-Choline Acetyltransferase antibody [EPR13024(B)]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

IHC image of Choline Acetyltransferase staining in a section of frozen normal human cerebral cortex performed on a Leica BOND^TM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab181023, 1/2000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

Intracellular Flow Cytometry analysis of SH-SY5Y cells labeling Choline Acetyltransferase using ab181023 (unpurified) at a 1/110 dilution (pink). Goat anti rabbit IgG (FITC) used as the secondary at a 1/150 dilution. Isotype control Rabbit monoclonal IgG (green). Cells were fixed in 2% paraformaldehyde.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebrum tissue sections labeling Choline Acetyltransferase with purified ab181023 at 1/2000 dilution (0.28 μg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

Immunocytochemistry/ Immunofluorescence analysis of U-87 MG (Human glioblastoma-astrocytoma epithelial cell) cells labeling Choline Acetyltransferase with purified ab181023 at 1/50 dilution (10 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

Immunofluorescent analysis of 4% paraformaldehyde fixed SH-SY5Y cells labeling Choline Acetyltransferase using ab181023 (unpurified) at a 1/100 dilution. A Goat anti rabbit IgG (Alexa Fluor®555) was used as the secondary at a 1/100 dilution.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Choline Acetyltransferase with purified ab181023 at 1/60 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IP

Unknown

Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

ab181023 (purified) at 1/30 dilution (2ug) immunoprecipitating Choline Acetyltransferase in Human fetal brain lysate. Human fetal brain lysate 10ug
Lane 2 (+) : ab181023 & Human fetal brain lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab181023 in Human fetal brain lysate
For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

Predicted band size: 82 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

Lysate from Human fetal brain (Lane 1) and negative control (Lane 2) were immunoprecipitated with ab181023 (unpurified) at a 1/70 dilution. A anti-rabbit IgG (HRP), specific to the non-reduced form of IgG at a 1/1500 dilution for the secondary. Blocking/ Dilution buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

Predicted band size: 82 kDa

false

• WB

Supplier Data

Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

Blocking/ Dilution buffer : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023) at 1/5000 dilution

Lane 1:

SH-SY5Y cell lysate at 20 µg

Lane 2:

Human fetal brain lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 82 kDa

false

• WB

Lab

Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (AB181023)

This antibody recognizes all Choline Acetyltransferase isoforms with the MW ranging from 70-82 KDa based on immunogen blast.

All lanes:

Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023) at 1/2000 dilution

Lane 1:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates at 20 µg

Lane 2:

Mouse brain lysates at 20 µg

Lane 3:

Rat brain lysates at 20 µg

Lane 4:

C6 (Rat glial tumor glial cell) whole cell lysates at 20 µg

Lane 5:

Human fetal brain lysates at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 82 kDa

Observed band size: 70-82 kDa

false