Overview

  • Product name
    Anti-Nocturnin antibody
  • Description
    Rabbit polyclonal to Nocturnin
  • Host species
    Rabbit
  • Specificity
    Although there is a high homology with mouse, this antibody also has the potential to cross react with another mouse protein of similar molecular weight therefore this antibody is not suitable for use in mouse.
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Rat, Cow
  • Immunogen

    A region within a synthetic peptide: RLYSALAKTL NSSAASQHPE YLVSPDPEHL EPIDPKELLE ECRAVLHTRP, corresponding to N terminal amino acids 73-122 of Human Nocturnin

  • Positive control
    • Human lung tissue (IHC-P), Daudi cell lysate (western blot).

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.09% Sodium azide
    Constituents: 2% Sucrose, PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab22850 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 a concentration of 1 - 2 µg/ml. Detects a band of approximately 40 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use at an assay dependent concentration.

Target

  • Relevance
    CCRN4L is highly similar to Nocturnin, a gene identified as a circadian clock regulated gene in Xenopus laevis. This protein and Nocturnin protein share similarity with the C terminal domain of a yeast transcription factor, carbon catabolite repression 4 (CCR4). The mRNA abundance of a similar gene in mouse has been shown to exhibit circadian rhythmicity, which suggests a role for this protein in clock function or as a circadian clock effector.
  • Database links
  • Alternative names
    • CCR4 carbon catabolite repression 4 like (S. cerevisiae) antibody
    • CCR4 protein homolog antibody
    • CCRN4L antibody
    • NOC antibody

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung tissue labelling Nocturnin with ab22850 at 4.0-8.0µg/ml. Positive staining seen in the bronchiole epithelium. Magnification: 400X.
  • Anti-Nocturnin antibody (ab22850) at 4 µg/ml + Daudi cell lysate

    Observed band size: 40 kDa
    why is the actual band size different from the predicted?



    Gel concentration: 12%
  • Paraffin-embedded Human lung tissue. Cells with positive label: epithelial cells of bronchiole (indicated by arrows). Antibody concentration: 4-8µg/ml. Magnification: 400x.

References

ab22850 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (3T3 cells (fibroblasts))
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
3T3 cells (fibroblasts)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 10 2015

Answer

Thank you for getting back to me with further information. I have read your previous questionnaire and the further details that you have provided me with. I find your enquiry most interesting. As I can see you are ONLY detecting the correctly sized and expressed Nocturin following immunoprecipitation. The western blots that you have performed are clearly detecting extraneous bands but not the FLAG tagged product when using this antibody. However, the Flag antibody detects the correctly sized Nocturin and demonstrates an enrichment following IP. The recognition of the FLAG tagged expressed Nocturin by western blot analysis is obviously the root of the problem. I ask under what conditions have you performed the detection. I ask this because we often find diluting the antibody in BSA with an overnight incubation at 4oC leads to heightened specificity by western blotting. I look forward to hearing from you.

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I have read your technical questionnaire and I have a few comments. You mention that this antibody has been detecting bands other than overexpressed murine and human nocturin proteins. Please can you tell me whether these bands could possibly be degradation products as I would recommend that the lysis buffer is supplemented with additional protease inhibitors. I would also appreciate your comments on whether these extraneous bands could be the result of truncated products from your expression construct. Please can you tell me whether you have eliminated this possibility by probing with an antibody against the FLAG tag. (This assumes that the FLAG is positioned at the N terminus; else FLAG will ONLY detect the full sized protein). In your questionnaire you mentioned that you thought that this antibody was detecting bands other than Nocturnin. Can you tell me whether there are bands detected in the negative controls? Finally I would greatly appreciate it if you could e-mail me an image of a representative blot as this will greatly assist me in understanding the pattern of extraneous bands that you have been detecting. I look forward to hearing from you.

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

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