Overview

  • Product name

  • Description

    Rabbit polyclonal to Nogo A
  • Host species

    Rabbit
  • Specificity

    ab62024 is specific for Nogo A and Nogo E.
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Raised against a 19 amino acid synthetic peptide from near the centre of Human Nogo A (GenBank accession no. NP_065393).

  • Positive control

    • Human brain tissue and mouse brain tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab62024 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
IHC-P
WB
  • Application notes
    ICC/IF: Use at a concentration of 1 µg/ml.
    IHC-P: Use at a concentration of 2.5 µg/ml.
    WB: Use at a concentration of 0.5 - 1.0 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 130 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function

      Developmental neurite growth regulatory factor with a role as a negative regulator of axon-axon adhesion and growth, and as a facilitator of neurite branching. Regulates neurite fasciculation, branching and extension in the developing nervous system. Involved in down-regulation of growth, stabilization of wiring and restriction of plasticity in the adult CNS. Regulates the radial migration of cortical neurons via an RTN4R-LINGO1 containing receptor complex (By similarity). Isoform 2 reduces the anti-apoptotic activity of Bcl-xl and Bcl-2. This is likely consecutive to their change in subcellular location, from the mitochondria to the endoplasmic reticulum, after binding and sequestration. Isoform 2 and isoform 3 inhibit BACE1 activity and amyloid precursor protein processing.
    • Tissue specificity

      Isoform 1 is specifically expressed in brain and testis and weakly in heart and skeletal muscle. Isoform 2 is widely expressed except for the liver. Isoform 3 is expressed in brain, skeletal muscle and adipocytes. Isoform 4 is testis-specific.
    • Sequence similarities

      Contains 1 reticulon domain.
    • Domain

      Three regions, residues 59-172, 544-725 and the loop 66 amino acids, between the two transmembrane domains, known as Nogo-66 loop, appear to be responsible for the inhibitory effect on neurite outgrowth and the spreading of neurons. This Nogo-66 loop, mediates also the binding of RTN4 to its receptor.
    • Cellular localization

      Endoplasmic reticulum membrane. Anchored to the membrane of the endoplasmic reticulum through 2 putative transmembrane domains.
    • Information by UniProt
    • Database links

    • Alternative names

      • ASY antibody
      • Foocen antibody
      • Human NogoA antibody
      • My043 protein antibody
      • Nbla00271 antibody
      • Nbla10545 antibody
      • neurite growth inhibitor 220 antibody
      • Neurite outgrowth inhibitor antibody
      • Neuroendocrine-specific protein antibody
      • Neuroendocrine-specific protein C homolog antibody
      • NI220/250 antibody
      • Nogo A antibody
      • NOGO antibody
      • Nogo protein antibody
      • NOGO-A antibody
      • Nogo-B antibody
      • Nogo-C antibody
      • NSP antibody
      • NSP-CL antibody
      • Reticulon 4 antibody
      • reticulon 5 antibody
      • Reticulon-4 antibody
      • Reticulon-5 antibody
      • RTN 4 antibody
      • RTN 4A antibody
      • RTN xL antibody
      • RTN-x antibody
      • Rtn4 antibody
      • RTN4-A antibody
      • RTN4-B1 antibody
      • RTN4-B2 antibody
      • RTN4-C antibody
      • RTN4_HUMAN antibody
      see all

    Images

    • Lane 1 : Anti-Nogo A antibody (ab62024) at 0.5 µg/ml
      Lane 2 : Anti-Nogo A antibody (ab62024) at 1 µg/ml

      All lanes : Human brain tissue lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : anti-rabbit IgG

      Predicted band size: 130 kDa
      Observed band size: 180 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 115 kDa, 155 kDa. We are unsure as to the identity of these extra bands.

    • Immunohistochemistry (Formalin-fixed paraffin embedded sections) of mouse brain tissue labeling Nogo A with Anti-Nogo A antibody (ab62024) at 5μg/ml.
    • ab62024, at 2.5µg/ml, staining Mouse Nogo A in brain tissue, by Immunohistochemistry
    • ICC/IF image of ab62024 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 (ab62024, 1µg/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.

    • Immunofluorescence of NogoA in Mouse Brain cells using ab62024 at 20 ug/ml.

    References

    This product has been referenced in:

    • Thau-Zuchman O  et al. Brain Phospholipid Precursors Administered Post-Injury Reduce Tissue Damage and Improve Neurological Outcome in Experimental Traumatic Brain Injury. J Neurotrauma 36:25-42 (2019). Read more (PubMed: 29768974) »
    • Mukherjee RN & Levy DL Reticulon 4a promotes exocytosis in mammalian cells. Mol Biol Cell 30:2349-2357 (2019). Read more (PubMed: 31318320) »
    See all 8 Publications for this product

    Customer reviews and Q&As

    1-5 of 5 Abreviews or Q&A

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa cell)
    Permeabilization
    Yes - 0.1-0.25% Triton X 100
    Specification
    HeLa cell
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Dr. Daniel Levy

    Verified customer

    Submitted May 23 2017

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa cell lysate)
    Gel Running Conditions
    Reduced Denaturing (5-20% gradient gel)
    Loading amount
    10 µg
    Treatment
    Benzonase nuclease for 5-10 min
    Specification
    HeLa cell lysate
    Blocking step
    LICOR Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0% · Temperature: RT°C

    Dr. Daniel Levy

    Verified customer

    Submitted May 23 2017

    Question
    Answer

    Thank you for contacting us yesterday to discuss the comments your reviewer had made.

    I have had a look into your enquiry and have come up with the following information:

    Nogo A is a potent neurite outgrowth inhibitor and plays a role both in the restriction of axonal regeneration after injury and in structural plasticity in the CNS of higher vertebrates.

    From the studies below, it seems that it is expressed in the oligodendrocytes and neurons

    Gil et al. J. Neuropathol. Exp. Neurol., 2006, 65(4), 433

    Wang et al. J. Neurosci., 2002, 22(12), 5505

    It seems that Nogo A shows an inhibitory effect on axonal regeneration when expressed in Schwann cells. It seems that Nogo A is not usually expressed in these cells:

    Pot et al. J. Cell Bio. Oct 14;159(1):29-35

    I'm afraid I still don't really understand why the reviewer has asked for this staining to be performed. Maybe you could go back to ask for further clarification from them?

    I hope this information has been of some help. If I can be of any further assistance, please do not hesitate to ask.

    Read More

    Question

    Here are the blots. The marker on the left, starting from the bottom is 20, 30, 40, 50, 60, 80, 100, 120 kDa.
     
    From: technical@abcam.com [mailto:technical@abcam.com]
    Sent: Thursday, January 26, 2012 4:08 PM
    To: Leitner, Dominique
    Subject: Reply from Abcam to your enquiry regarding ab62024, ab65800 [CCE3459244]


     



    https://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header


    Dear Dominique
    We have an answer to your inquiry:

    Thank you for contacting Abcam. 
     

    It appears as if you may have recognized an isoform of Nogo. The immunogen sequence used corresponds to aa117601192 of Nogo A. However, this sequence is also present in each of the four known isoforms of Nogo. Would you be able to send me an image of you Western blot? 
     

    I look forward to your response. Please let me know if you have any questions.
     

     

    Help us improve our service.
    https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3459244



    Your original inquiry to Abcam:
     

     
     

     
     

     
     

     
     
     
     

     
     

     
     

     
     

     
     






     
     

    I have a question about an antibody. I have used NogoA ab65800 antibody to do a western blot with rat brain lysate. I saw a band at about 58kDa. I block for one hour in 5% milk TBST, incubate in primary overnight, and incubate in secondary for one hour at room temperature. It was developed by ECL. The predicted size is listed as 130kDa and is shown with a rat brain lysate around here. What would you recommend? Can this antibody be replaced with another NogoA antibody (ab62024)?
     

     
     

    Thanks,
     

     
     

    Dominique Leitner

    GSA Vice President

    GWIS President

    Ph.D. Candidate, Neuroscience

    Dept of Neurosurgery, H110

    Penn State University College of Medicine

    Milton S. Hershey Medical Center

    500 University Drive

    Hershey, PA 17033

    Phone: 717-531-5203

    Email: dleitner@hmc.psu.edu
     

     
     







     
     
    Best regards,
    Keith

    Keith Beadle
    Scientific Support Specialist
    Abcam Inc.
    www.abcam.com
     
    Abcam Customer Services and Scientific Support Team
    https://www.abcam.com/index.html?pageconfig=technical&utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Body
    [CCE3459244]






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    Read More
    Answer

    Thank you for that additional information. This has helped clarify the issues that you have been experiencing.

    I am sorry that this antibody has not worked as it should have. The image that you have provided us shows definitively that this is a random stain and not a Nogo isoform. I would like to replace the product with ab62024 as you have asked but will need to confirm the original purchase information with you.

    Could you please send me the order number or PO number with which the original product was purchased. After I recieve this information I will process a free of charge replacement for you.

    Please let me know if you have any questions.

    Read More

    Answer

    Thank you for contacting Abcam.

    It appears as if you may have recognized an isoform of Nogo. The immunogen sequence used corresponds to aa117601192 of Nogo A. However, this sequence is also present in each of the four known isoforms of Nogo. Would you be able to send me an image of you Western blot?

    I look forward to your response. Please let me know if you have any questions.

    Read More

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