Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (AB45689)

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (AB45689)

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).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (AB45689)

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.

• IP

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Immunoprecipitation - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (AB45689)

ab45689 immunoprecipitating Calmodulin. 10 μg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

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

All lanes:

Immunoprecipitation - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)

Observed band size: 16 kDa

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• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (AB45689)

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 Fluorr^® 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)

• WB

Lab

Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (AB45689)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689) at 1/10000 dilution

Lane 1:

RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate at 15 µg

Lane 2:

C6 (rat glioma) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 17 kDa

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• WB

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Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (AB45689)

All lanes:

Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689) at 1/5000 dilution

All lanes:

NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate at 10 µg

Predicted band size: 17 kDa

Observed band size: 16 kDa

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• WB

PubMed

Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (AB45689)

ΔSRCR peripheral blood monocyte-derived macrophages (PMMs) still function as hemoglobin-haptoglobin (Hb-Hp) scavengers.

Porcine PMMs were incubated for 24 hours in the presence of 100 μg/mol Hb-Hp then lysed with reducing SDS sample buffer and HO-1 protein expression analyzed by western blot.

For full details please see paper.

All lanes:

Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)

Predicted band size: 17 kDa

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Burkard et al PLoS Pathog. 2017 Feb 23;13(2):e1006206. doi: 10.1371/journal.ppat.1006206. eCollection 2017 Feb. Fig 8. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• WB

Lab

Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (AB45689)

All lanes:

Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689) at 1/10000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma) whole cell lysate at 15 µg

Lane 2:

HCT 116 (human colorectal carcinoma) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 17 kDa,40 kDa

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