Product Info

Source:
Mouse
Application:
WB 1:500-2000, IHC 1:200-1000, FCM 1:200-400, ELISA 1:10000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Clonality:
Monoclonal [AFB14950]
Specificity:
PCNA antibody detects endogenous levels of total PCNA.
RRID:
AB_2843434
Cite Format: Affinity Biosciences Cat# BF0704, RRID:AB_2843434.
Conjugate:
Unconjugated.
Purification:
Affinity-chromatography.
Storage:
Mouse IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

ATLD2; cb16; Cyclin; DNA polymerase delta auxiliary protein; etID36690.10; fa28e03; fb36g03; HGCN8729; MGC8367; Mutagen-sensitive 209 protein; OTTHUMP00000030189; OTTHUMP00000030190; PCNA; Pcna/cyclin; PCNA_HUMAN; PCNAR; Polymerase delta accessory protein; Proliferating cell nuclear antigen; wu:fa28e03; wu:fb36g03;

Immunogens

Immunogen:

Purified recombinant fragment of human PCNA (AA: 53-196 ) expressed in E. Coli.

Uniprot:
Gene(ID):
Description:
PCNA This protein is an auxiliary protein of DNA polymerase delta and is involved in the control of eukaryotic DNA replication by increasing the polymerase's processibility during elongation of the leading strand. Belongs to the PCNA family. Homotrimer. Forms a complex with activator 1 heteropentamer in the presence of ATP.
Sequence:
MFEARLVQGSILKKVLEALKDLINEACWDISSSGVNLQSMDSSHVSLVQLTLRSEGFDTYRCDRNLAMGVNLTSMSKILKCAGNEDIITLRAEDNADTLALVFEAPNQEKVSDYEMKLMDLDVEQLGIPEQEYSCVVKMPSGEFARICRDLSHIGDAVVISCAKDGVKFSASGELGNGNIKLSQTSNVDKEEEAVTIEMNEPVQLTFALRYLNFFTKATPLSSTVTLSMSADVPLVVEYKIADMGHLKYYLAPKIEDEEGS

PTMs - P12004 As Substrate

Site PTM Type Enzyme
R5 Methylation
S10 Phosphorylation
K13 Acetylation
K13 Ubiquitination
K14 Acetylation
K14 Ubiquitination
Y60 Phosphorylation
R61 Methylation
K77 Acetylation
K77 Ubiquitination
K80 Acetylation
K80 Ubiquitination
C81 S-Nitrosylation
K110 Ubiquitination
K117 Ubiquitination
Y133 Phosphorylation
S134 Phosphorylation
C135 S-Nitrosylation
K138 Ubiquitination
R146 Methylation
S152 Phosphorylation
S161 Phosphorylation
C162 S-Nitrosylation
K164 Acetylation
K164 Sumoylation
K164 Ubiquitination
K168 Ubiquitination
K181 Ubiquitination
T185 Phosphorylation
K190 Acetylation
R210 Methylation
Y211 Phosphorylation P00533 (EGFR)
T216 Phosphorylation
K217 Acetylation
S228 Phosphorylation
K248 Acetylation
K248 Ubiquitination
Y250 Phosphorylation
K254 Sumoylation
K254 Ubiquitination
S261 Phosphorylation

Research Backgrounds

Function:

Auxiliary protein of DNA polymerase delta and is involved in the control of eukaryotic DNA replication by increasing the polymerase's processibility during elongation of the leading strand. Induces a robust stimulatory effect on the 3'-5' exonuclease and 3'-phosphodiesterase, but not apurinic-apyrimidinic (AP) endonuclease, APEX2 activities. Has to be loaded onto DNA in order to be able to stimulate APEX2. Plays a key role in DNA damage response (DDR) by being conveniently positioned at the replication fork to coordinate DNA replication with DNA repair and DNA damage tolerance pathways. Acts as a loading platform to recruit DDR proteins that allow completion of DNA replication after DNA damage and promote postreplication repair: Monoubiquitinated PCNA leads to recruitment of translesion (TLS) polymerases, while 'Lys-63'-linked polyubiquitination of PCNA is involved in error-free pathway and employs recombination mechanisms to synthesize across the lesion.

PTMs:

Phosphorylated. Phosphorylation at Tyr-211 by EGFR stabilizes chromatin-associated PCNA.

Acetylated by CREBBP and p300/EP300; preferentially acetylated by CREBBP on Lys-80, Lys-13 and Lys-14 and on Lys-77 by p300/EP300 upon loading on chromatin in response to UV irradiation. Lysine acetylation disrupts association with chromatin, hence promoting PCNA ubiquitination and proteasomal degradation in response to UV damage in a CREBBP- and EP300-dependent manner. Acetylation disrupts interaction with NUDT15 and promotes degradation.

Ubiquitinated. Following DNA damage, can be either monoubiquitinated to stimulate direct bypass of DNA lesions by specialized DNA polymerases or polyubiquitinated to promote recombination-dependent DNA synthesis across DNA lesions by template switching mechanisms. Following induction of replication stress, monoubiquitinated by the UBE2B-RAD18 complex on Lys-164, leading to recruit translesion (TLS) polymerases, which are able to synthesize across DNA lesions in a potentially error-prone manner. An error-free pathway also exists and requires non-canonical polyubiquitination on Lys-164 through 'Lys-63' linkage of ubiquitin moieties by the E2 complex UBE2N-UBE2V2 and the E3 ligases, HLTF, RNF8 and SHPRH. This error-free pathway, also known as template switching, employs recombination mechanisms to synthesize across the lesion, using as a template the undamaged, newly synthesized strand of the sister chromatid. Monoubiquitination at Lys-164 also takes place in undamaged proliferating cells, and is mediated by the DCX(DTL) complex, leading to enhance PCNA-dependent translesion DNA synthesis. Sumoylated during S phase.

Methylated on glutamate residues by ARMT1/C6orf211.

Subcellular Location:

Nucleus.
Note: Colocalizes with CREBBP, EP300 and POLD1 to sites of DNA damage (PubMed:24939902). Forms nuclear foci representing sites of ongoing DNA replication and vary in morphology and number during S phase. Together with APEX2, is redistributed in discrete nuclear foci in presence of oxidative DNA damaging agents.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Subunit Structure:

Homotrimer. Interacts with p300/EP300; the interaction occurs on chromatin in UV-irradiated damaged cells. Interacts with CREBBP (via transactivation domain and C-terminus); the interaction occurs on chromatin in UV-irradiated damaged cells. Directly interacts with POLD1, POLD3 and POLD4 subunits of the DNA polymerase delta complex, POLD3 being the major interacting partner; the interaction with POLD3 is inhibited by CDKN1A/p21(CIP1). Forms a complex with activator 1 heteropentamer in the presence of ATP. Interacts with EXO1, POLH, POLK, DNMT1, ERCC5, FEN1, CDC6 and POLDIP2. Interacts with APEX2; this interaction is triggered by reactive oxygen species and increased by misincorporation of uracil in nuclear DNA. Forms a ternary complex with DNTTIP2 and core histone. Interacts with KCTD10 and PPP1R15A (By similarity). Directly interacts with BAZ1B. Interacts with HLTF and SHPRH. Interacts with NUDT15; this interaction is disrupted in response to UV irradiation and acetylation. Interacts with CDKN1A/p21(CIP1) and CDT1; interacts via their PIP-box which also recruits the DCX(DTL) complex. The interaction with CDKN1A inhibits POLD3 binding. Interacts with DDX11. Interacts with EGFR; positively regulates PCNA. Interacts with PARPBP. Interacts (when ubiquitinated) with SPRTN; leading to enhance RAD18-mediated PCNA ubiquitination. Interacts (when polyubiquitinated) with ZRANB3. Interacts with SMARCAD1. Interacts with CDKN1C. Interacts with PCLAF (via PIP-box). Interacts with RTEL1 (via PIP-box); the interaction is direct and essential for the suppression of telomere fragility. Interacts with FAM111A (via PIP-box); the interaction is direct and required for PCNA loading on chromatin binding. Interacts with LIG1. Interacts with SETMAR. Interacts with ANKRD17. Interacts with FBXO18/FBH1 (via PIP-box); the interaction recruits the DCX(DTL) complex and promotes ubiquitination and degradation of FBXO18/FBH1. Interacts with POLN. Interacts with SDE2 (via PIP-box); the interaction is direct and prevents ultraviolet light induced monoubiquitination. Component of the replisome complex composed of at least DONSON, MCM2, MCM7, PCNA and TICRR; interaction at least with PCNA occurs during DNA replication. Interacts with MAPK15; the interaction is chromatin binding dependent and prevents MDM2-mediated PCNA destruction by inhibiting the association of PCNA with MDM2. Interacts with PARP10 (via PIP-box). Interacts with DDI2. Interacts with HMCES (via PIP-box).

(Microbial infection) Interacts with herpes virus 8 protein LANA1.

Family&Domains:

Belongs to the PCNA family.

Research Fields

· Cellular Processes > Cell growth and death > Cell cycle.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Tight junction.   (View pathway)

· Genetic Information Processing > Replication and repair > DNA replication.

· Genetic Information Processing > Replication and repair > Base excision repair.

· Genetic Information Processing > Replication and repair > Nucleotide excision repair.

· Genetic Information Processing > Replication and repair > Mismatch repair.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

References

1). Combination of resolvin E1 and lipoxin A4 promotes the resolution of pulpitis by inhibiting NF-κB activation through upregulating sirtuin 7 in dental pulp fibroblasts. CELL PROLIFERATION, 2022 (PubMed: 35411569) [IF=8.5]

Application: WB    Species: rat    Sample: Dental pulp fibroblasts

FIGURE 2 |Effects of the RvE1 and LXA4 combination on NF-κB modification(E) Expression of p65 protein in both cytoplasm and nucleus detected by western blotting, normalized to β-actin (in cytoplasm) or PCNA (in nucleus).

2). Role of Annexin A2 isoform 2 on the aggregative growth of dermal papillae cells. BIOSCIENCE REPORTS, 2018 (PubMed: 30341243) [IF=4.0]

Application: WB    Species: human    Sample: passage 3 DPC

Figure 4.| Effect of RNA interference mediated inhibition of Annexin A2 isoform2 expression on biological function of passage 3 DPC.(d)Western blot results showed the expression level of PCNA was suppressed in passage 3 DPC.

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