Immunocytochemistry/ Immunofluorescence - ALDH1A1 antibody - Neuronal Marker (ab23375)

ICC/IF image of ab23375 stained rat PC12 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab23375, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - ALDH1A1 antibody - Neuronal Marker (ab23375)

IHC image of ALDH1A1 staining in rat brain FFPE section, performed on a Bond^TM 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 20 mins. The section was then incubated with ab23375, 1µg/ml, for 8 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.

- ALDH1A1 antibody - Neuronal Marker (ab23375)

ab23375 staining ALDH1A1 in Mouse hypothalamic tanycytes by IHC-FrFl (Immunohistochemistry Free-floating). 40 µm free-floating sections were incubated with primary antibody (1/300) for 18 hours at 4ºC in blocking solution. An Alexa Fluor^® 555-conjugated Donkey polyclonal to rabbit IgG (1/300) was used as secondary antibody.

This image is courtesy of an anonymous Abreview

Western blot - ALDH1A1 antibody - Neuronal Marker (ab23375)

All lanes : Anti-ALDH1A1 antibody - Neuronal Marker (ab23375) at 1 µg/ml

Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 2 : Liver (Mouse) Tissue Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 54 kDa
Observed band size : 54 kDa
Additional bands at : 95 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 90 seconds

Western blot - ALDH1A1 antibody - Neuronal Marker (ab23375)

All lanes : Anti-ALDH1A1 antibody - Neuronal Marker (ab23375) at 1 µg/ml

Lane 1 : Brain (Mouse) Tissue Lysate
Lane 2 : Liver (Mouse) Tissue Lysate
Lane 3 : Liver (Rat) Tissue Lysate
Lane 4 : E14 Mouse Embryo Brain Tissue Lysate

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 54 kDa
Observed band size : 51,54 kDa (why is the actual band size different from the predicted?)


Exposure time : 90 seconds

Western blot - Anti-ALDH1A1 antibody - Neuronal Marker (ab23375)

Anti-ALDH1A1 antibody - Neuronal Marker (ab23375) at 1 µg/ml + ALDH1A1 protein (Human) (ab89491) at 0.1 µg

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Exposure time : 1 minute