The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 67 kDa (predicted molecular weight: 67 kDa).
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. PubMed: 23556002
Use a concentration of 5 µg/ml.
FunctionRequired for the relief of p53-dependent checkpoint mediated cell cycle arrest. Binds to and dephosphorylates 'Ser-15' of TP53 and 'Ser-345' of CHEK1 which contributes to the functional inactivation of these proteins.
Sequence similaritiesBelongs to the PP2C family. Contains 1 PP2C-like domain.
protein phosphatase 1D magnesium-dependent delta isoform antibody
Protein phosphatase 2C delta isoform antibody
Protein phosphatase 2C isoform delta antibody
Protein phosphatase magnesium dependent 1 delta antibody
Protein phosphatase magnesium-dependent 1 delta antibody
Protein phosphatase Mg2+/Mn2+ dependent 1D antibody
Protein phosphatase Wip1 antibody
Wild type p53 induced phosphatase 1 antibody
WIP 1 antibody
Anti-PPM1D antibody images
Western blot - PPM1D antibody (ab31270)
All lanes : Anti-PPM1D antibody (ab31270) at 1 µg/ml
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 67 kDa Observed band size : 67 kDa
Exposure time : 20 minutes
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPM1D antibody (ab31270)Image from Li et al., PLoS One, 8(3): e60775, Figure 2a. doi:10.1371/journal.pone.0060775
Immunohistochemical staining using ab31720 in paraffin-embedded HCC and non-cancerous liver tissues, 5 µm sections, at a dilution of 1/1000. (a) Negative ppm1d staining in normal liver tissue. (b) Negative staining of HCC tissue without ab31720. (c) Ppm1d-positive non-cancerous liver tissue. (d) High expression of ppm1d in HCC tissue. Antigen retrieval was performed in a microwave with 0.1 M sodium citrate buffer (pH 6.0). The secondary antibody was a HRP conjugated compact polymer system and the samples were counter stained with DAB. These images were taken on a Keyence BZ-8000 digital microscope at a magnification of x200 and the scale bar represents 100 µm.
IHC image of ab31270 staining in Human breast adenocarcinoma 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 ab31270, 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.
ICC/IF image of ab31270 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab31270, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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-PPM1D antibody (ab31270)
This product has been referenced in:
Li GB et al. Protein phosphatase magnesium-dependent 1d (PPM1D) mRNA expression is a prognosis marker for hepatocellular carcinoma. PLoS One8:e60775 (2013).
Read more (PubMed: 23556002) »
Zajkowicz A et al. Nutlin-3a, an MDM2 antagonist and p53 activator, helps to preserve the replicative potential of cancer cells treated with a genotoxic dose of resveratrol. Mol Biol Rep40:5013-26 (2013).
Read more (PubMed: 23666059) »