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

9 product images

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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/

  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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  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

  Secondary

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

  Predicted band size: 17 kDa, 40 kDa

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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) 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).

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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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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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  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.

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

  Predicted band size: 17 kDa

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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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  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) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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.

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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)