The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1/2000 - 1/10000. Detects a band of approximately 171 kDa (predicted molecular weight: 185 kDa).
Use at 1-4 µg/mg of lysate.
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.
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
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
SCN circadian oscillatory protein antibody
Suprachiasmatic nucleus circadian oscillatory protein antibody
Suprachiasmatic nucleus circadian protein antibody
Anti-PHLPP antibody images
Western blot - PHLPP antibody (ab71972)
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
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 One9:e98858 (2014).
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 Res19:4662-72 (2013).
Read more (PubMed: 23846336) »