Anti-Calbindin antibody [EP3478] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

This data was developed using ab108404, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human cerebellum staining of Calbindin with ab108404 at a 1/1000 dilution, DGKZ/DGK-zeta with ab239081 at 1/5000 dilution, and GABA A Receptor alpha 6 with ab300069 at a 1/500 dilution, followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-GABA A Receptor alpha 6 (gray; Opal™690), anti-DGKZ/DGK-zeta (green; Opal™520) and anti-Calbindin (magenta; Opal™570) on human cerebellum.
Panel B : anti-Calbindin stained on all layers of cerebellum.
Panel C : anti-DGKZ/DGK-zeta stained on Purkinje cells.
Panel D : anti-GABA A Receptor alpha 6 stained on the granular layer.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300069, ab239081, and ab108404 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

ab108404 showing negative staining in Normal colon tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Calbindin with purified ab108404 at 1/150 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). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

ab108404, at 1/100 dilution, staining Calbindin in paraffin-embedded Human brain tissue by Immunohistochemistry.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

Immunofluorescent analysis was performed on 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse hippocampal primary neural/glia cells labelling Calbindin with ab108404 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody (ab150077) at dilution of 1/1000 (green).

Anti-MAP2 antibody [HM-2] (ab11267) at 1/1000 dilution was used as a counterstain, with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed as the secondary antibody (ab150120) at dilution of 1/1000 (red).

Negative control 1 : Anti-Calbindin (ab108404) at 1/250 dilution with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed as the secondary antibody (ab150120) at dilution of 1/1000.

Negative control 2 : Anti-MAP2 antibody [HM-2] (ab11267) at 1/200 dilution with Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody (ab150077) at dilution of 1/1000.

The nuclear counterstain is DAPI (blue).

Confocal image showing positive staining in subsets of mouse hippocampal primary neuron.

Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

ab108404 showing negative staining in Normal liver tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

ab108404 showing negative staining in Normal heart tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EP3478] - BSA and Azide free (AB233018)

ab108404 showing positive staining in Normal kidney tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108404).