Overview

  • Product nameAnti-PHLPP antibody
    See all PHLPP primary antibodies
  • Description
    Rabbit polyclonal to PHLPP
  • Tested applicationsSuitable for: ICC/IF, IHC-P, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Chimpanzee, Rhesus monkey, Gorilla, Opossum, Orangutan
  • Immunogen

    Synthetic peptide corresponding to a region between residue 1150 and the C-terminus (residue 1205) of human PHLPP (NP_919431.1)

  • Positive control
    • Whole cell lysate from HeLa, 293T or NIH3T3 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab71972 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 a concentration of 1 µg/ml.
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/2000 - 1/10000. Detects a band of approximately 171 kDa (predicted molecular weight: 185 kDa).
IP Use at 1-4 µg/mg of lysate.

Target

  • FunctionProtein phosphatase that mediates dephosphorylation of 'Ser-473' of AKT1, 'Ser-660' of PRKCB isoform beta-II and 'Ser-657' of PRKCA. AKT1 regulates the balance between cell survival and apoptosis through a cascade that primarily alters the function of transcription factors that regulate pro- and antiapoptotic genes. Dephosphorylation of 'Ser-473' of AKT1 triggers apoptosis and suppression of tumor growth. Controls the phosphorylation of AKT2 and AKT3 more efficiently than that of AKT1. Dephosphorylation of PRKCA and PRKCB leads to their destabilization and degradation. Inhibits cancer cell proliferation and may act as a tumor suppressor. May act as a negative regulator of K-Ras signaling in membrane rafts.
  • Tissue specificityIn colorectal cancer tissue, expression is highest in the surface epithelium of normal colonic mucosa adjacent to the cancer tissue but is largely excluded from the crypt bases. Expression is lost or significantly decreased in 78% of tested tumors (at protein level). Ubiquitously expressed in non-cancerous tissues.
  • Sequence similaritiesContains 21 LRR (leucine-rich) repeats.
    Contains 1 PH domain.
    Contains 1 PP2C-like domain.
  • DomainThe PH domain is required for interaction with PRKCB and its dephosphorylation.
  • Cellular localizationCytoplasm. Membrane. Nucleus. In colorectal cancer tissue, expression is concentrated at the lateral membrane of epithelial cells.
  • Information by UniProt
  • Database links
  • Alternative names
    • hSCOP antibody
    • KIAA0606 antibody
    • PH domain and leucine rich repeat protein phosphatase antibody
    • PH domain leucine rich repeat containing protein phosphatase antibody
    • PH domain leucine rich repeat protein phosphatase antibody
    • PH domain leucine-rich repeat-containing protein phosphatase 1 antibody
    • PH domain-containing family E member 1 antibody
    • PHLP1_HUMAN antibody
    • PHLPP antibody
    • Phlpp1 antibody
    • Pleckstrin homology domain containing family E (with leucine rich repeats) member 1 antibody
    • Pleckstrin homology domain containing family E protein 1 antibody
    • Pleckstrin homology domain-containing family E member 1 antibody
    • PLEKHE 1 protein antibody
    • PLEKHE1 antibody
    • SCN circadian oscillatory protein antibody
    • SCOP antibody
    • Suprachiasmatic nucleus circadian oscillatory protein antibody
    • Suprachiasmatic nucleus circadian protein antibody
    see all

Anti-PHLPP antibody images

  • All lanes : Anti-PHLPP antibody (ab71972) at 0.04 µg/ml

    Lane 1 : HeLa whole cell lysate at 50 µg
    Lane 2 : HeLa whole cell lysate at 15 µg
    Lane 3 : HeLa whole cell lysate at 5 µg
    Lane 4 : 293T whole cell lysate at 50 µg
    Lane 5 : NIH3T3 whole cell lysate at 50 µg


    Predicted band size : 185 kDa
    Observed band size : 171 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 238 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 10 seconds
  • Detection of PHLPP by Western Blot of Immunoprecipitate.
    ab71972 at 1µg/ml staining PHLPP in HeLa whole cell lysate immunoprecipitated using ab71972 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane).
    Detection: chemiluminescence with exposure time of 30 seconds.
  • ab71972 (2µg/ml) staining PHLPP in human cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of the cytoplasm.
    Inset panel depicts negative control Rabbit IgG at 2µg/ml.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • ICC/IF image of ab71972 stained Hek293 cells. The cells were 4% PFA 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 (ab71972, 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.

References for Anti-PHLPP antibody (ab71972)

This product has been referenced in:
  • Endo H  et al. Dormancy of cancer cells with suppression of AKT activity contributes to survival in chronic hypoxia. PLoS One 9:e98858 (2014). WB ; Human . Read more (PubMed: 24905002) »
  • Liao WT  et al. microRNA-224 promotes cell proliferation and tumor growth in human colorectal cancer by repressing PHLPP1 and PHLPP2. Clin Cancer Res 19:4662-72 (2013). WB ; Human . Read more (PubMed: 23846336) »

See all 3 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (LNCaP - prostate cancer cells)
Gel Running Conditions Reduced Denaturing (8% SDS-PAGE)
Loading amount 20 µg
Specification LNCaP - prostate cancer cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Dec 03 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (LNCaP - prostate cancer cells)
Permeabilization Yes - 0.2% Triton X-100
Specification LNCaP - prostate cancer cells
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

Submitted Nov 27 2015

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

Dr. Robert Kupp

Verified customer

Submitted Aug 18 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (FIBROBLASTS)
Loading amount 15 µg
Specification FIBROBLASTS
Gel Running Conditions Reduced Denaturing (8% GEL)
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Imogen Butcher

Verified customer

Submitted Jul 08 2010

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