Anti-Caspr antibody (ab34151)

Anti-Caspr antibody images

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  Western blot - Caspr antibody (ab34151)
  Anti-Caspr antibody (ab34151) at 1/250 dilution + Brain (Rat) Whole Cell Lysate - normal tissue at 10 µg
  
  Secondary
  IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size : 156 kDa
  Observed band size : 180 kDa (why is the actual band size different from the predicted?)
  Additional bands at : 58 kDa. We are unsure as to the identity of these extra bands.Caspr contains a number of potential glycosylation sites so it is thought that this is the reason it runs at 180kDa.
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  Immunocytochemistry/ Immunofluorescence - Caspr antibody (ab34151)
  ICC/IF image of ab34151 stained SHSY5Y cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab34151, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).
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  Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Caspr antibody (ab34151)This image is courtesy of an abreview submitted by Sophie Pezet, ESPCI, France
  IHC-FoFR image of ab34151 stained sections of mouse brain (30 um). The tissues were from perfused fixed animals perfused with 4% PFA and postfixed 2h in the same fixative. They were cryoprotected in 30% sucrose and cut using a cryostat.

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  Immunoprecipitation - Anti-Caspr antibody (ab34151)
  Caspr was immunoprecipitated using 0.5mg Rat Brain whole tissue lysate, 5µg of Rabbit polyclonal to Caspr 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 ab34151.
  Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
  Band: 180kDa: Caspr.
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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Caspr antibody (ab34151)Image courtesy of an anonymous Abreview.
  ab34151 staining Caspr in murine brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde. Samples were blocked with 20% serum for 1 hour at 20°C followed by incubation with the primary antibody at a 1/300 dilution for 12 hours at 20°C. An HRP-conjugated goat anti-rabbit polyclonal was used as the secondary antibody at a 1/200 dilution.

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