Overview

  • Product nameAnti-Cyclin A antibodySee all Cyclin A2 primary antibodies ...
  • Description
    Rabbit polyclonal to Cyclin A
  • Tested applicationsWB, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Hamster
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Mouse Cyclin A.

    (Peptide available as ab95425.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: F9; MEF1; K562.

Properties

Applications

Our Abpromise guarantee covers the use of ab87359 in the following tested applications.

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

Application Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 47 kDa).Can be blocked with Mouse Cyclin A peptide (ab95425).
ICC/IF Use a concentration of 5 µg/ml.

Target

Anti-Cyclin A antibody images

  • All lanes : Anti-Cyclin A antibody (ab87359) at 1 µg/ml

    Lane 1 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : K562 (Human erythromyeloblastoid leukemia 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 : 47 kDa
    Observed band size : 55 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 minutes
  • ICC/IF image of ab87359 stained HepG2 cells. The cells were 100% Methanol fixed (5 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 (ab87359, 5µg/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) HepG2 cells at 5µg/ml.
  • ab87359 staining cyclin A in DU145 cells treated with lovastatin (ab120614), by ICC/IF. Decrease in cyclin A expression correlates with increased concentration of lovastatin, as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120614 (lovastatin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab87359(5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

References for Anti-Cyclin A antibody (ab87359)

ab87359 has not yet been referenced specifically in any publications.

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