Recombinant Anti-Calbindin antibody [EP3478] (ab108404)

All lanes : Anti-Calbindin antibody [EP3478] (ab108404) at 1/2000 dilution (Purified)

Lane 1 : Human fetal brain lysates
Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
Lane 3 : Human kidney lysates
Lane 4 : Mouse brain lysates
Lane 5 : Mouse kidney lysates
Lane 6 : Rat brain lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 30 kDa
Observed band size: 30 kDa

Immunohistochemistry (Frozen) analysis of mouse cerebellum tissue sections labeling Calbindin with purified ab108404 at 1/500 dilution (3.0 µg/ml). The tissue section was fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. DAPI (blue) was used as nuclear counterstain.
Secondary antibody only control: PBS instead of the primary antibody

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebrum tissue sections labeling Calbindin with purified ab108404 at 1/1000 dilution (1.49 µg/ml). Perform 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.

All lanes : Anti-Calbindin antibody [EP3478] (ab108404) at 1/1000 dilution

Lane 1 : Fetal brain lysate
Lane 2 : Fetal cerebellum lysate
Lane 3 : Fetal hypothalamus lysate
Lane 4 : Fetal pituitary lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 30 kDa

Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Calbindin with purified ab108404 at 1/150 dilution (10 µg/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).

Immunohistochemistry (Frozen) analysis of rat cerebellum tissue sections labeling Calbindin with purified ab108404 at 1/500 dilution (3.0 µg/ml). The tissue section was fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. DAPI (blue) was used as nuclear counterstain.
Secondary antibody only control: PBS instead of the primary antibody

Immunocytochemistry/ Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Calbindin with purified ab108404 at 1/150 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) dilution. 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.

Immunohistochemical analysis of mouse frozen cerebellum tissue sections with ab108404 labelling Calbindin at 1/500. Sections were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).  The image shows specific staining on Purkinje cells of mouse cerebellum.

Secondary antibody only was used as negative control and is shown in the inset.

ab108404 showing positive staining in Normal kidney tissue.

Immunocytochemistry/Immunofluorescence analysis of Neuro-2a (mouse neuroblastoma) cells labelling Calbindin with ab108404 at 1/100. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at dilution of 1/200. Nuclei were counterstained with DAPI (blue).

Confocal image showing nuclear and cytoplasmic staining on Neuro2a cultured with 2% FBS.

ab108404, at 1/100 dilution, staining Calbindin in paraffin-embedded Human brain tissue by Immunohistochemistry.

ab108404 showing negative staining in Normal heart tissue.

ab108404 showing negative staining in Normal colon tissue.

ab108404 showing negative staining in Normal liver tissue.