Protein phosphatase 4C consists of a catalytic subunit PPP4C and a regulatory subunit. PPP4R1 and belongs to the PPP phosphatase family, PP X subfamily. Protein phosphatases are involved in determining the phosphorylation state of many regulatory proteins. PPP4C is involved in many processes such as microtubule organization at centrosomes, maturation of spliceosomal snRNPs, apoptosis, tumor necrosis factor (TNF)-alpha signaling, activation of c-Jun N-terminal kinase MAPK8, regulation of histone acetylation, DNA damage checkpoint signaling, NF-kappa-B activation and cell migration. The PPP4SLCPPP4R1 PP4 complex may play a role in dephosphorylation and regulation of HDAC3. The PPP4SLCPPP4R2-PPP4R3A PP4 complex specifically dephosphorylates H2AFX phosphorylated on Ser-140 (gamma-H2AFX) generated during DNA replication and required for DNA DSB repair. Dephosphorylates NDEL1 at CDK1 phosphorylation sites and negatively regulates CDK1 activity in interphase.
Function:
Protein phosphatase that is involved in many processes such as microtubule organization at centrosomes, maturation of spliceosomal snRNPs, apoptosis, DNA repair, tumor necrosis factor (TNF)-alpha signaling, activation of c-Jun N-terminal kinase MAPK8, regulation of histone acetylation, DNA damage checkpoint signaling, NF-kappa-B activation and cell migration. The PPP4SLCPPP4R1 PP4 complex may play a role in dephosphorylation and regulation of HDAC3. The PPP4SLCPPP4R2-PPP4R3A PP4 complex specifically dephosphorylates H2AFX phosphorylated on Ser-140 (gamma-H2AFX) generated during DNA replication and required for DNA double strand break repair. Dephosphorylates NDEL1 at CDK1 phosphorylation sites and negatively regulates CDK1 activity in interphase. In response to DNA damage, catalyzes RPA2 dephosphorylation, an essential step for DNA repair since it allows the efficient RPA2-mediated recruitment of RAD51 to chromatin.
Subunit:
Serine/threonine-protein phosphatase 4 (PP4) occurs in different assemblies of the catalytic and one or more regulatory subunits. Component of the PP4 complexes PPP4SLCPPP4R1, PPP4SLCPPP4R2, PPP4SLCPPP4R2-PPP4R3A, PPP4SLCPPP4R2-PPP4R3B and PPP4SLCPPP4R4. The PPP4SLCPPP4R2 complex appears to be a tetramer composed of 2 molecules of PPP4C and 2 molecules of PPP4R2. Interacts with REL, NFKB1/p50 and RELA. Interacts with SMN1 AND GEMIN4. Interacts with IRS4 (phosphorylated). Interacts with SMEK1/PPP4R3A; the interaction requires PP4R2. Interacts with HDAC3.
Subcellular Location:
Cytoplasm. Nucleus. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome.
Similarity:
Belongs to the PPP phosphatase family. PP-4 (PP-X) subfamily.
SWISS:
P60510
Gene ID:
5531
Database links:
Entrez Gene: 5531 Human
Entrez Gene: 56420 Mouse
Entrez Gene: 100009163 Rabbit
Entrez Gene: 171366 Rat
Omim: 602035 Human
SwissProt: P60510 Human
SwissProt: P97470 Mouse
SwissProt: P11084 Rabbit
SwissProt: Q5BJ92 Rat
Unigene: 534338 Human
Unigene: 41998 Mouse
Unigene: 9173 Rat
激酶和磷酸酶（Kinases and Phosphatases）
蛋白磷酸酶-4(又称PP4或PPX)是蛋白磷酸酶2A(PP2A)家族的重要成员之一；
PP-2Ac参与细胞周期,生长和分化及多细胞信号通路的调节，也用于肿瘤方面的研究。
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Sample:
Kidney (Mouse) Lysate at 40 ug
Primary: Anti-PPP4C (SL0490R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 34 kD
Observed band size: 34 kD
Tissue/cell: human gastric carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-PPP4C Polyclonal Antibody, Unconjugated(SL0490R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-PPP4C Polyclonal Antibody, Unconjugated(SL0490R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (PPP4C) polyclonal Antibody, Unconjugated (SL0490R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Blank control: HepG2.
Primary Antibody (green line): Rabbit Anti-PPP4C antibody (SL0490R)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Blank control: HepG2.
Primary Antibody (green line): Rabbit Anti-PPP4C antibody (SL0490R)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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