Anti-CNPase antibody [mAbcam 44289] (ab44289)

Overview

• Product nameAnti-CNPase antibody [mAbcam 44289]See all CNPase primary antibodies ...
• Description
  Mouse monoclonal [mAbcam 44289] to CNPase
• Tested applicationsIP, Flow Cyt, IHC-FoFr, IHC-P, WB more details
• Species reactivity
  Reacts with: Mouse, Rat, Dog, Human
  Predicted to work with: Orangutan
• Immunogen

  Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human CNPase.

  Read Abcam's proprietary immunogen policy

  (Peptide available as ab19424.)

• Positive controlThis antibody gave a positive signal in the following Tissue Lysates: Human Brain, Mouse Brain, Mouse Spinal Cord, Rat Brain, Rat Spinal Cord
• General notes

  We can conjugate this antibody to FITC for you (please see ab150257 for details).

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: PBS, pH 7.4
• Concentration information loading...
• PurityIgG fraction
• Clonality Monoclonal
• Clone numbermAbcam 44289
• MyelomaSp2/0
• IsotypeIgG1
• Research Areas
  • Tags & Cell Markers
  • Cell Type Markers
  • Neuroscience Markers
  • Glial

  • Neuroscience
  • Cell Type Marker
  • Glia marker
  • Oligodendrocyte marker

  • Neuroscience
  • Neurology process
  • Neurodegenerative disease
  • Other

Applications

Target

• Sequence similaritiesBelongs to the cyclic nucleotide phosphodiesterase family.
• Cellular localizationMembrane. Melanosome. Firmly bound to membrane structures of brain white matter. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.

Target information above from: UniProt accession P09543 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 607694 Dog
  • Entrez Gene: 1267 Human
  • Entrez Gene: 12799 Mouse
  • Entrez Gene: 25275 Rat
  • Omim: 123830 Human
  • SwissProt: P09543 Human
  • SwissProt: P16330 Mouse
  • SwissProt: P13233 Rat

  • Unigene: 273621 Human
  • Unigene: 15711 Mouse
  • Unigene: 31762 Rat

  see all

• Alternative names
  2'' antibody2'3' cyclic nucleotide 3' phosphodiesterase antibody2'3'-cyclic-nucleotide 3'-phosphodiesterase antibody

  3''-cyclic-nucleotide 3''-phosphodiesterase antibodyCN37_HUMAN antibodyCNP 1 antibodyCNP antibodyCNP1 antibodyCNPase antibody

  see all

Anti-CNPase antibody [mAbcam 44289] images

• 

  Western blot - CNPase antibody (ab44289)
  Anti-CNPase antibody [mAbcam 44289] (ab44289) at 5 µg/ml + Brain (Human) Tissue Lysate - adult normal tissue (ab29466) at 10 µg
  
  Secondary
  Rabbit polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size : 46,48 kDa
  Observed band size : 46,48 kDa
  
• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [mAbcam 44289] (ab44289)Carl Hobbs, King`s College London, United Kingdom
  IHC-P image of CNPase staining on human brain section using ab44289 (1:100). The paraffin embedded section was subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked using 1% BSA at 21°C for 10 min. ab44289 was incubated at 21°C for 16 hours. The secondary antibody was goat polyclonal anti-mouse IgG conjugated to biotin (1:200)

  See Abreview

• 

  Western blot - CNPase antibody [mAbcam 44289] (ab44289)
  All lanes : Anti-CNPase antibody [mAbcam 44289] (ab44289) at 1 µg/ml
  
  Lane 1 : Brain (Mouse) Tissue Lysate
  Lane 2 : Spinal Cord (Mouse) Tissue Lysate
  Lane 3 : Brain (Rat) Tissue Lysate
  Lane 4 : Spinal Cord (Rat) Tissue Lysate
  Lane 5 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 46,48 kDa
  Observed band size : 46,48 kDa
  Additional bands at : 42 kDa,67 kDa. We are unsure as to the identity of these extra bands.
  
  Exposure time : 3 minutes
• 

  Flow Cytometry-Anti-CNPase antibody [mAbcam 44289](ab44289)
  Overlay histogram showing SHSY-5Y cells stained with ab44289 (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 (ab44289, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse 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 SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  
  
• 

  Immunoprecipitation - Anti-CNPase antibody [mAbcam 44289] (ab44289)
  CNPase was immunoprecipitated using 0.5mg Rat Brain whole tissue lysate, 5µg of Mouse monoclonal to CNPase and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
  The antibody was incubated under agitation with Protein G beads for 10min, Rat Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
  Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab44289.
  Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
  Band: 48,46kDa: CNPase.
• 

  Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-CNPase antibody [mAbcam 44289] (ab44289)Sophie Pezet, Laboratoire de Neurobiologie, France
  IHC-FoFr image of CNPase staining on rat dentate gyrus sections using ab44289(1:300). The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

  See Abreview