Overview

  • Product name

    Anti-INPPL1/SHIP-2 antibody
    See all INPPL1/SHIP-2 primary antibodies
  • Description

    Rabbit polyclonal to INPPL1/SHIP-2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Chimpanzee, Rhesus monkey, Gorilla, Chinese hamster
  • Immunogen

    Synthetic peptide corresponding to Human INPPL1/SHIP-2 aa 100-150.
    Database link: NP_001558.2

  • Positive control

  • General notes

    Previously labelled as INPPL1. 

Properties

Applications

Our Abpromise guarantee covers the use of ab70267 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 Use at an assay dependent concentration. PubMed: 21757001
WB 1/2000 - 1/20000. Detects a band of approximately 139 kDa (predicted molecular weight: 139 kDa).
IP Use at 1-4 µg/mg of lysate.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    Phosphatidylinositol (PtdIns) phosphatase that specifically hydrolyzes the 5-phosphate of phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P3) to produce PtdIns(3,4)P2, thereby negatively regulating the PI3K (phosphoinositide 3-kinase) pathways. Plays a central role in regulation of PI3K-dependent insulin signaling, although the precise molecular mechanisms and signaling pathways remain unclear. While overexpression reduces both insulin-stimulated MAP kinase and Akt activation, its absence does not affect insulin signaling or GLUT4 trafficking. Confers resistance to dietary obesity. May act by regulating AKT2, but not AKT1, phosphorylation at the plasma membrane. Part of a signaling pathway that regulates actin cytoskeleton remodeling. Required for the maintenance and dynamic remodeling of actin structures as well as in endocytosis, having a major impact on ligand-induced EGFR internalization and degradation. Participates in regulation of cortical and submembraneous actin by hydrolyzing PtdIns(3,4,5)P3 thereby regulating membrane ruffling. Regulates cell adhesion and cell spreading. Required for HGF-mediated lamellipodium formation, cell scattering and spreading. Acts as a negative regulator of EPHA2 receptor endocytosis by inhibiting via PI3K-dependent Rac1 activation. Acts as a regulator of neuritogenesis by regulating PtdIns(3,4,5)P3 level and is required to form an initial protrusive pattern, and later, maintain proper neurite outgrowth. Acts as a negative regulator of the FC-gamma-RIIA receptor (FCGR2A). Mediates signaling from the FC-gamma-RIIB receptor (FCGR2B), playing a central role in terminating signal transduction from activating immune/hematopoietic cell receptor systems. Involved in EGF signaling pathway. Upon stimulation by EGF, it is recruited by EGFR and dephosphorylates PtdIns(3,4,5)P3. Plays a negative role in regulating the PI3K-PKB pathway, possibly by inhibiting PKB activity. Down-regulates Fc-gamma-R-mediated phagocytosis in macrophages independently of INPP5D/SHIP1. In macrophages, down-regulates NF-kappa-B-dependent gene transcription by regulating macrophage colony-stimulating factor (M-CSF)-induced signaling. May also hydrolyze PtdIns(1,3,4,5)P4, and could thus affect the levels of the higher inositol polyphosphates like InsP6.
  • Tissue specificity

    Widely expressed, most prominently in skeletal muscle, heart and brain. Present in platelets. Expressed in transformed myeloid cells and in primary macrophages, but not in peripheral blood monocytes.
  • Involvement in disease

    Defects in INPPL1 may be a cause of susceptibility to type 2 diabetes mellitus non-insulin dependent (NIDDM) [MIM:125853].
    Note=Genetic variations in INPPL1 may be a cause of susceptibility to metabolic syndrome. Metabolic syndrome is characterized by diabetes, insulin resistance, hypertension, and hypertriglyceridemia is absent.
  • Sequence similarities

    Belongs to the inositol 1,4,5-trisphosphate 5-phosphatase family.
    Contains 1 SAM (sterile alpha motif) domain.
    Contains 1 SH2 domain.
  • Domain

    The SH2 domain interacts with tyrosine phosphorylated forms of proteins such as SHC1 or FCGR2A. It also mediates the interaction with p130Cas/BCAR1.
    The NPXY sequence motif found in many tyrosine-phosphorylated proteins is required for the specific binding of the PID domain.
  • Post-translational
    modifications

    Tyrosine phosphorylated by the members of the SRC family after exposure to a diverse array of extracellular stimuli such as insulin, growth factors such as EGF or PDGF, chemokines, integrin ligands and hypertonic and oxidative stress. May be phosphorylated upon IgG receptor FCGR2B-binding. Phosphorylated at Tyr-986 following cell attachment and spreading. Phosphorylated at Tyr-1162 following EGF signaling pathway stimulation. Phosphorylated at Thr-958 in response to PDGF.
  • Cellular localization

    Cytoplasm > cytosol. Cytoplasm > cytoskeleton > actin patch. Membrane. Translocates to membrane ruffles when activated, translocation is probably due to different mechanisms depending on the stimulus and cell type. Partly translocated via its SH2 domain which mediates interaction with tyrosine phosphorylated receptors such as the FC-gamma-RIIB receptor (FCGR2B). Tyrosine phosphorylation may also participate in membrane localization. Insulin specifically stimulates its redistribution from the cytosol to the plasma membrane. Recruited to the membrane following M-CSF stimulation.
  • Information by UniProt
  • Database links

  • Alternative names

    • 4 antibody
    • 5-trisphosphate 5-phosphatase 2 antibody
    • 51C protein antibody
    • EC 3.1.3.n1 antibody
    • inositol polyphosphate phosphatase like 1 antibody
    • Inositol polyphosphate phosphatase like protein 1 antibody
    • Inositol polyphosphate phosphatase-like protein 1 antibody
    • INPPL-1 antibody
    • INPPL1 antibody
    • OPSMD antibody
    • Phosphatidylinositol 3 antibody
    • Phosphatidylinositol 3,4,5 trisphosphate 5 phosphatase 2 antibody
    • Phosphatidylinositol-3 antibody
    • Protein 51C antibody
    • SH2 domain containing inositol 5' phosphatase 2 antibody
    • SH2 domain-containing inositol 5''-phosphatase 2 antibody
    • SH2 domain-containing inositol phosphatase 2 antibody
    • SHIP-2 antibody
    • SHIP2 antibody
    • SHIP2_HUMAN antibody
    see all

Images

  • Immunocytochemistry/Immunofluorescence analysis of human GIST882 cells labelling INPPL1/SHIP-2 (top) with ab70267. Bottom - Hoechst 33342.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling INPPL1/SHIP-2 with ab70267 at 1/1000 (1µg/ml). Detection: DAB.

  • Anti-INPPL1/SHIP-2 antibody (ab70267) at 0.1 µg/ml + HeLa whole cell lysate at 50 µg

    Developed using the ECL technique.

    Predicted band size: 139 kDa
    Observed band size: 139 kDa


    Exposure time: 10 seconds
  • Immunoprecipitation of HeLa whole cell lysate (1mg) using ab70267 at 3 µg/mg. 1/4 of the immunoprecipitate was loaded on to a gel and analysed by Western blot using ab70267 at 0.1 µg/ml. Blot was developed using chemiluminescence with an exposure time of 30 seconds.
  • IHC image of ab70267 staining in Human breast cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab70267, 5µg/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.

References

This product has been referenced in:

  • Deneubourg L  et al. Abnormal elevated PTEN expression in the mouse antrum of a model of GIST Kit(K641E/K641E). Cell Signal 23:1857-68 (2011). ICC/IF ; Mouse . Read more (PubMed: 21757001) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunoprecipitation
Sample
Rat Cell lysate - whole cell (L6 myotubes)
Total protein in input
1000 µg
Immuno-precipitation step
Protein G
Specification
L6 myotubes

Abcam user community

Verified customer

Submitted Apr 03 2019

Answer

Thank you for contacting us.

Every experiement has different optimal antibody concentrations, and therefore proper working dilutions for every antibody must be optimized for the system in which it is being used.

However, I suggest that you start with 5ug/ml (1:200 dilution) of antibody as a starting concentration for IF to see how good the staining is.

For the titration, I would suggest you use a range of concentrations ranging from 1-5ug/ml. For example:

5 ug/ml (1:200)
2.5 ug/ml (1:400)
1.6 ug/ml (1:600)
1.25 ug/ml (1:800)
1ug/ml (1:1000)

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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