Overview

  • Product nameAnti-CRMP5 antibody
    See all CRMP5 primary antibodies
  • Description
    Rabbit polyclonal to CRMP5
  • Tested applicationsSuitable for: WB, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 450 - 550 of Human CRMP5.

    (Peptide available as ab89373.)

  • Positive control
    • This antibody gave a positive signal in human brain and human spinal cord tissue lysates and in the following whole cell lysates: SHSY-5Y; SK N BE; SHSY-5Y.

Properties

Applications

Our Abpromise guarantee covers the use of ab79008 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
WB Use at an assay dependent concentration. Detects a band of approximately 70 kDa (predicted molecular weight: 61 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IP Use a concentration of 5 µg/ml.

Target

Anti-CRMP5 antibody images

  • CRMP5 was immunoprecipitated using 0.5mg SHSY5Y whole cell extract, 5µg of Rabbit polyclonal to CRMP5 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, SHSY5Y whole cell extract 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°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab79008.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 70kDa; CRMP5, non specific band - 55kDa: We are unsure as to the identity of this extra band.

  • All lanes : Anti-CRMP5 antibody (ab79008) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human spinal cord tissue lysate - total protein (ab29188)
    Lane 3 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 4 : SK N BE (Human neuroblastoma) Whole Cell Lysate
    Lane 5 : SHSY-5Y (Human neuroblastoma 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 : 61 kDa
    Observed band size : 70 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 110 kDa,55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes
    CRMP5 contains a number of potential phosphorylation sites (SwissProt data) which may explain it running at a higher molecular weight than predicted.
  • ICC/IF image of ab79008 stained SHSY5Y cells. The cells were 4% formaldehyde 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 (ab79008, 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 100% methanol fixed (5 min) SHSY5Y cells at 5µg/ml.

References for Anti-CRMP5 antibody (ab79008)

ab79008 has not yet been referenced specifically in any publications.

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Zebrafish Tissue sections (retinal ganglion cells)
Permeabilization Yes - 0.1% Triton X
Specification retinal ganglion cells
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative Paraformaldehyde
Username

Dr. Ryan MacDonald

Verified customer

Submitted Mar 12 2014

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