Overview

  • Product nameAnti-PKC antibody [M110]
    See all PKC primary antibodies
  • Description
    Mouse monoclonal [M110] to PKC
  • SpecificityThis antibody detects 80-82 kDa proteins corresponding to the molecular mass of PKC alpha, PKC beta and PKC gamma on SDS-PAGE immunoblots of neonatal rat brain lysates. Similar results were observed in human and mouse lysates. Immunoprecipitation experiments with various PKC isoforms demonstrated that this antibody detects PKC alpha, PKC beta and PKC gamma, but not other PKC isoforms expressed in rat brain lysate.
  • Tested applicationsICC/IF, IHC-P, IP, ELISA, WB, ICC, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment, corresponding to C terminal amino acids 499-697 of Human PKC

  • Positive control
    • Human, mouse, rat brain lysate.
  • General notesDo not aliquot.

Properties

Applications

Our Abpromise guarantee covers the use of ab23511 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
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP Use at an assay dependent dilution.
ELISA 1/2000.
WB 1/1000. Detects a band of approximately 80 kDa.
ICC 1/100.
Flow Cyt 1/100.

Target

Anti-PKC antibody [M110] images

  • Lane 1 : Anti-PKC antibody [M110] (ab23511) at 1/250 dilution
    Lane 2 : Anti-PKC antibody [M110] (ab23511) at 1/500 dilution
    Lane 3 : Anti-PKC antibody [M110] (ab23511) at 1/1000 dilution

    Lane 1 : rat brain lysate
    Lane 2 : rat brain lysate
    Lane 3 : rat brain lysate


    Observed band size : 81 kDa (why is the actual band size different from the predicted?)
    Membrane was incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1 hour at room temperature.
  • ICC/IF image of ab23511 stained PC12 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 (ab23511, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

  • IHC image of ab23511 staining in human normal skin 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 ab23511, 10µg/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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Overlay histogram showing Jurkat cells stained with ab23511 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab23511, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-PKC antibody [M110] (ab23511)

This product has been referenced in:
  • Boncoeur E  et al. Modulation of epithelial sodium channel activity by lipopolysaccharide in alveolar type II cells: involvement of purinergic signaling. Am J Physiol Lung Cell Mol Physiol 298:L417-26 (2010). WB ; Rat . Read more (PubMed: 20008115) »

See 1 Publication for this product

Product Wall

Thank you for your enquiry. Immunoprecipitation experiments with various PKC isoforms demonstrated that this antibody detects PKC alpha, PKC beta and PKC gamma, but not other PKC isoforms. I hope this information helps, please do not hesitate to ...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"