Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP799Y] to Calmodulin 1/2/3 - C-terminal
  • Suitable for: WB, IHC-P, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal
    See all Calmodulin 1/2/3 primary antibodies

  • Description

    Rabbit monoclonal [EP799Y] to Calmodulin 1/2/3 - C-terminal

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Calmodulin 1/2/3 aa 100 to the C-terminus. The exact sequence is proprietary.

  • Positive control

    • WB: C6, RAW264.7, NIH/3T3, HeLa, HCT 116 cell lysate. IHC-P: Human urinary bladder carcinoma and testis tissues. Flow Cyt: MCF7 and NIH/3T3 cells. IP: Human skeletal muscle tissue lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

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

    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.

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

    Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 2 : HCT 116 (human colorectal carcinoma) whole cell lysate

    Blocking peptides at 15 µg per lane.

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

    Predicted band size: 17 kDa



    Blocking and Diluting buffer: 5% NFDM/TBST

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)
    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)
    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.

  • Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)
    Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)
    All lanes : 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
    Lane 2 : C6 (rat glioma) whole cell lysate

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 17 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)
    Western blot - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)
    Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689) at 1/5000 dilution + NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate at 10 µg

    Predicted band size: 17 kDa
    Observed band size: 16 kDa
    why is the actual band size different from the predicted?

  • Flow Cytometry - Anti-Calmodulin 1/2/3 antibody [EP799Y] - C-terminal (ab45689)
    Flow Cytometry - 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.

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

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

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

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