Anti-TGN46 antibody - Golgi Marker

• ICC

AbReview13480****

Immunocytochemistry - Anti-TGN46 antibody - Golgi Marker (AB16059)

Paraformaldehyde-fixed, Triton-X 100-permeabilised mouse keratinocytes stained for TGN46 at 1/200 dilution in ICC.

• ICC

Lab

Immunocytochemistry - Anti-TGN46 antibody - Golgi Marker (AB16059)

ab16059 staining TGN46 in MEF1 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab16059 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGN46 antibody - Golgi Marker (AB16059)

IHC image of ab16059 staining in mouse e14 whole foetus formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16059, 5µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TGN46 antibody - Golgi Marker (AB16059)

IHC image of TGN46 staining in a section of formalin-fixed paraffin-embedded normal mouse E14 embryo performed on a Leica BOND™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab16059, 1μg/ml, 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.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-TGN46 antibody - Golgi Marker (AB16059)

Immunocytochemistry-immunofluorescence using Anti-TGN46 antibody - Golgi Marker, ab16059. Publication image from Frosch, M. P. et al., 2015, Nat Commun, 26458742. Legend direct from paper.

Neuronal uptake of PBS-soluble HMW tau derived from human AD brain.(a,b) Primary neurons were incubated with AD or control brain extracts (cases were matched for age and postmortem interval (Supplementary Table S1)) and immunostained at day 2 (a). (b) Quantification of fluorescence intensity of human tau staining. One-way ANOVA and a subsequent Tukey-Kramer test. (c,d) Tau uptake (c) and seeding activity (d) assay in HEK-tau-biosensor cells. (Mann–Whitney U-test) (e) Subcellular localization of human tau taken up by neurons (PBS-3,000g, 500 ng ml−1 human tau). (f) Neuron-to-neuron transfer of tau in a 3-chamber microfluidic device. AD brain extract (PBS-3,000g, 500 ng ml−1 human tau) was added to the 1st chamber. Human tau positive neurons were detected in both the 1st and 2nd chamber at day 7 (arrow). (g,h) Quantification of total-tau (g) and phospho-tau (h) levels in AD and control brain extract (ELISA). Unpaired t-test. (i) Brain extracts were immunoblotted with phospho-tau specific antibodies recognizing different epitopes. Representative immunoblot and quantification of phospho-tau levels at each epitope. Unpaired t-test. (j,k) SEC analysis of PBS-soluble tau from AD and control brain. (j) Representative graph of total tau levels (ELISA) in SEC-separated samples. Small peaks for HMW fractions were detected in both groups (right panel). (k) Mean total tau levels of HMW SEC fractions. (l) Tau uptake from each SEC fraction (5 or 500 ng ml−1 human tau) by primary neurons. (m) Phospho-tau levels in each SEC fraction (ELISA). Unpaired t-test. Scale bar, 25 µm. *P<0.05, **P<0.01.