Anti-Calbindin antibody - Neuronal Marker (ab49899)

Western blot - Calbindin antibody (ab49899)

All lanes : Anti-Calbindin antibody - Neuronal Marker (ab49899) at 1 µg/ml

Lane 1 : Kidney (Human) Tissue Lysate - adult normal tissue (ab30203)
Lane 2 : Kidney (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

Performed under reducing conditions.

Predicted band size : 30 kDa
Observed band size : 30 kDa
Additional bands at : 150 kDa. We are unsure as to the identity of these extra bands.

Immunohistochemistry (PFA perfusion fixed frozen sections) - Calbindin antibody - Neuronal Marker (ab49899)

Immunohistochemistical detection of Calbindin using antibody ab49899 on PFA perfusion-fixed frozen rat brain sections (30um). Antigen retrieval step: None Primary Antibody diluted at 1/300 and incubated for 18 hours @ 20°C in PBS + 0.3 % Triton X100. Secondary Antibody: Goat anti-rabbit Alexa Fluor® 488 (1/1000). ab49899 was used on sections of rat brain in free floating IHC; expected staining was observed on rat brain. The submitted images show staining obtained in the cerebral cortex (A,B). The images were taken using the objective X5 (A) and X40 (B) and show the neuronal profiles (arrows) and neuronal processes.

Sophie Pezet, CNRS, Paris, France

Immunocytochemistry/ Immunofluorescence - Calbindin antibody - Neuronal Marker (ab49899)

ICC/IF image of ab49899 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab49899, 5µg/ml) overnight at +4ºC. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Calbindin antibody - Neuronal Marker (ab49899)

IHC image of Calbindin staining in rat brain formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab49899, 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.