Anti-Calmodulin 1/2/3 antibody [EP799Y] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - BSA and Azide free (AB214793)

Overlay histogram showing MCF7 (Human breast adenocarcinoma cell line) cells stained with ab45689 (red line).

The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab45689, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C.

Isotype control antibody (black line) was rabbit monoclonal IgG (0.5μg/1x10⁶ cells) used under the same conditions.

Acquisition of >5,000 events was performed.

This antibody gave a decreased signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - BSA and Azide free (AB214793)

ab45689 at a 1 : 250 dilution staining Calmodulin in human urinary bladder carcinoma tissue by immunohistochemistry in paraffin embedded tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - BSA and Azide free (AB214793)

ab45689 staining Calmodulin in human testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control : PBS in place of primary antibody (inset).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

• IP

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Immunoprecipitation - Anti-Calmodulin 1/2/3 antibody [EP799Y] - BSA and Azide free (AB214793)

ab45689 immunoprecipitating Calmodulin. 10 μg of cell lysate was incubated with primary antibody at a dilution of 1/20. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1 : Human skeletal muscle lysate (10 μg)
Lane 2 : Human skeletal muscle lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab45689 in human skeletal muscle lysate

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

All lanes:

Immunoprecipitation - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (<a href='/en-us/products/primary-antibodies/calmodulin-1-2-3-antibody-ep799y-c-terminal-ab45689'>ab45689</a>)

Observed band size: 16 kDa

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• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - BSA and Azide free (AB214793)

ab45689 staining Calmodulin in NIH/3T3 (Mouse embryo fibroblast cell line) cells by intracellular flow cytometry.

Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype 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 (ab45689).