Overview

  • Product name

    Anti-NIPA (phospho S354) antibody
    See all NIPA primary antibodies
  • Description

    Rabbit polyclonal to NIPA (phospho S354)
  • Host species

    Rabbit
  • Specificity

    Detects NIPA only when phosphorylated at serine 354.
  • Tested applications

    Suitable for: ELISA, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic phosphopeptide derived from human NIPA around the phosphorylation site of serine 354 (T-R-SP-W-D).

  • Positive control

    • IHC-P: FFPE human testis tissue sections.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride

    Without Mg2+ and Ca2+
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Purified from rabbit antiserum by affinity chromatography using epitope specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab63557 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
ELISA 1/20000.
WB 1/500 - 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    Essential component of an SCF-type E3 ligase complex, SCF(NIPA), a complex that controls mitotic entry by mediating ubiquitination and subsequent degradation of cyclin B1 (CCNB1). Its cell-cycle-dependent phosphorylation regulates the assembly of the SCF(NIPA) complex, restricting CCNB1 ubiquitination activity to interphase. Its inactivation results in nuclear accumulation of CCNB1 in interphase and premature mitotic entry. May have an antiapoptotic role in NPM-ALK-mediated signaling events.
  • Tissue specificity

    Widely expressed. Highly expressed in heart, skeletal muscle and testis. Expressed in brain, placenta, lung, kidney, liver, pancreas, spleen, thymus, prostate, ovary small intestine and colon. Weakly or not expressed in leukocytes.
  • Pathway

    Protein modification; protein ubiquitination.
  • Sequence similarities

    Contains 1 C3HC-type zinc finger.
  • Developmental stage

    Weakly expressed in G0/G1 phases, abundant during S and G2/M phases, and strongly decreases thereafter.
  • Domain

    The F-box-like region is required for the interaction with SKP1.
  • Post-translational
    modifications

    Phosphorylated. Phosphorylated on Ser residues at G2/M phase, but not during S and G0 phases. May also be weakly phosphorylated on Tyr residues. Ser-354 phosphorylation, a major site during the course of cell-cycle-dedendent phosphorylation, results in its dissociation from the SCF(NIPA) complex, thereby preventing CCNB1 degradation leading to mitotic entry.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • hNIPA antibody
    • NIPA_HUMAN antibody
    • Nuclear interacting partner of ALK antibody
    • Nuclear interacting partner of anaplastic lymphoma kinase antibody
    • Nuclear-interacting partner of ALK antibody
    • Nuclear-interacting partner of anaplastic lymphoma kinase antibody
    • zc3hc1 antibody
    • Zinc finger C3HC type containing 1 antibody
    • Zinc finger C3HC-type protein 1 antibody
    see all

Images

  • IHC image of NIPA staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab63557, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-NIPA (phospho S354) antibody (ab63557) at 1/500 dilution

    Lane 1 : extracts from COS7 cells, treated
    with HU (2nM, 24hours)
    Lane 2 : extracts from COS7 cells, treated
    with HU (2nM, 24hours) with immunizing phosphopeptide at 10 µg

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 55 kDa
    Observed band size: 55 kDa
    Additional bands at: 30 kDa, 75 kDa. We are unsure as to the identity of these extra bands.

References

This product has been referenced in:

  • Zhao M  et al. Methylene blue exerts a neuroprotective effect against traumatic brain injury by promoting autophagy and inhibiting microglial activation. Mol Med Rep 13:13-20 (2016). WB ; Mouse . Read more (PubMed: 26572258) »
  • Illert AL  et al. Extracellular signal-regulated kinase 2 (ERK2) mediates phosphorylation and inactivation of nuclear interaction partner of anaplastic lymphoma kinase (NIPA) at G2/M. J Biol Chem 287:37997-8005 (2012). WB . Read more (PubMed: 22955283) »
See all 2 Publications for this product

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