Anti-Caldesmon antibody (ab68878)

Overview

  • Product nameAnti-Caldesmon antibody
    See all Caldesmon primary antibodies
  • Description
    Rabbit polyclonal to Caldesmon
  • Tested applicationsSuitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Cow, Chimpanzee, Rhesus monkey
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 750 to the C-terminus of Human Caldesmon.

    (Peptide available as ab86635.)

  • Positive control
    • This antibody gave a positive signal in SK N SH Whole Cell Lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab68878 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 78 kDa (predicted molecular weight: 93 kDa).
ICC/IF Use a concentration of 1 µg/ml.

Target

  • FunctionActin- and myosin-binding protein implicated in the regulation of actomyosin interactions in smooth muscle and nonmuscle cells (could act as a bridge between myosin and actin filaments). Stimulates actin binding of tropomyosin which increases the stabilization of actin filament structure. In muscle tissues, inhibits the actomyosin ATPase by binding to F-actin. This inhibition is attenuated by calcium-calmodulin and is potentiated by tropomyosin. Interacts with actin, myosin, two molecules of tropomyosin and with calmodulin. Also play an essential role during cellular mitosis and receptor capping.
  • Tissue specificityHigh-molecular-weight caldesmon (isoform 1) is predominantly expressed in smooth muscles, whereas low-molecular-weight caldesmon (isoforms 2, 3, 4 and 5) are widely distributed in non-muscle tissues and cells. Not expressed in skeletal muscle or heart.
  • Sequence similaritiesBelongs to the caldesmon family.
  • DomainThe N-terminal part seems to be a myosin/calmodulin-binding domain, and the C-terminal a tropomyosin/actin/calmodulin-binding domain. These two domains are separated by a central helical region in the smooth-muscle form.
  • Post-translational
    modifications
    In non-muscle cells, phosphorylation by CDK1 during mitosis causes caldesmon to dissociate from microfilaments. Phosphorylation reduces caldesmon binding to actin, myosin, and calmodulin as well as its inhibition of actomyosin ATPase activity. Phosphorylation also occurs in both quiescent and dividing smooth muscle cells with similar effects on the interaction with actin and calmodulin and on microfilaments reorganization.
  • Cellular localizationCytoplasm > cytoskeleton. Cytoplasm > myofibril. On thin filaments in smooth muscle and on stress fibers in fibroblasts (nonmuscle).
  • Information by UniProt
  • Database links
  • Alternative names
    • CAD antibody
    • CALD 1 antibody
    • CALD1 antibody
    • CALD1_HUMAN antibody
    • Caldesmon 1 antibody
    • Caldesmon 1 Isoform 1 antibody
    • Caldesmon 1 Isoform 2 antibody
    • Caldesmon 1 Isoform 3 antibody
    • Caldesmon 1 Isoform 4 antibody
    • Caldesmon 1 Isoform 5 antibody
    • Caldesmon antibody
    • Caldesmon1 antibody
    • CDM antibody
    • H CAD antibody
    • HCAD antibody
    • L CAD antibody
    • LCAD antibody
    • MGC21352 antibody
    • NAG22 antibody
    see all

Anti-Caldesmon antibody images

  • ICC/IF image of ab68878 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab68878, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa, HepG2, Hek293 and MCF7 cells at 5µg/ml.
  • All lanes : Anti-Caldesmon antibody (ab68878) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : SK N SH (Human neuroblastoma) Whole Cell Lysate
    Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 93 kDa
    Observed band size : 78 kDa (why is the actual band size different from the predicted?)
  • IHC image of Caldesmon staining in normal human kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab68878 at 5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

References for Anti-Caldesmon antibody (ab68878)

ab68878 has not yet been referenced specifically in any publications.

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I am assuming the species customer of tissue sections is human - antibodeis are asfollows;

The best anti annexin a1 antibody is ab47661;http://www.abcam.com/Annexin-A1-antibody-ab47661.html

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