• Western blot analysis of extracts of various cell lines, using POLD3 antibody (STJ29386) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) (STJS000856) at 1:10000 dilution. Lysates/proteins: 25 Mu g per lane. Blocking buffer: 3% non-fat dry milk in TBST. Detection: ECL Enhanced Kit. Exposure time: 60s.
  • Immunohistochemistry analysis of paraffin-embedded human colon carcinoma using POLD3 rabbit polyclonal antibody (STJ29386) at dilution of 1:300 (40x lens). Perform microwave antigen retrieval with 10 mM PBS buffer pH 7. 2 before commencing with immunohistochemistry staining protocol.
  • Immunofluorescence analysis of GFP-RNF168 transgenic U2OS cells using POLD3 antibody (STJ29386). Green:GFP-RNF168 fusion protein expression for DNA damage marker. Blue: DAPI for nuclear staining. RNF168 (GFP) can be used to mark cells damaged by UV-A laser for they always gather around DNA damage region.
  • Immunofluorescence analysis of U2OS cells using POLD3 antibody (STJ29386).

Anti-POLD3 antibody (197-466) (STJ29386)

SKU:
STJ29386

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Host: Rabbit
Applications: WB/IHC/IF
Reactivity: Human/Mouse
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit polyclonal antibody anti-POLD3 (197-466) is suitable for use in Western Blot, Immunohistochemistry and Immunofluorescence research applications.
Clonality: Polyclonal
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.02% Sodium Azide, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:500-1:2000
IF/ICC 1:50-1:100
IHC-P 1:100-1:500
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: POLD3
Gene ID: 10714
Uniprot ID: DPOD3_HUMAN
Immunogen Region: 197-466
Immunogen: Recombinant fusion protein containing a sequence corresponding to amino acids 197-466 of human POLD3 (NP_006582.1).
Immunogen Sequence: ASKAAAKTQETNKETKTEAK EVTNASAAGNKAPGKGNMMS NFFGKAAMNKFKVNLDSEQA VKEEKIVEQPTVSVTEPKLA TPAGLKKSSKKAEPVKVLQK EKKRGKRVALSDDETKETEN MRKKRRRIKLPESDSSEDEV FPDSPGAYEAESPSPPPPPS PPLEPVPKTEPEPPSVKSSS GENKRKRKRVLKSKTYLDGE GCIVTEKVYESESCTDSEEE LNMKTSSVHRPPAMTVKKE
Post Translational Modifications Ubiquitinated, but not targeted to the proteasome. Sumoylated. Sumoylation with SUMO3 may be predominant. Phosphorylation at Ser-458 is catalyzed in vitro by PKA. It is thought to decrease the affinity for PCNA and Pol-delta4 processivity. Can also be phosphorylated in vitro by CDK1-cyclin-A complex, as well as CDK2-cyclin-A and CDK2-cyclin-E complexes. PCNA interferes with CDK-cyclin phosphorylation.
Function Accessory component of both the DNA polymerase delta complex and the DNA polymerase zeta complex. As a component of the trimeric and tetrameric DNA polymerase delta complexes (Pol-delta3 and Pol-delta4, respectively), plays a role in high fidelity genome replication, including in lagging strand synthesis, and repair. Required for optimal Pol-delta activity. Stabilizes the Pol-delta complex and plays a major role in Pol-delta stimulation by PCNA. Pol-delta3 and Pol-delta4 are characterized by the absence or the presence of POLD4. They exhibit differences in catalytic activity. Most notably, Pol-delta3 shows higher proofreading activity than Pol-delta4. Although both Pol-delta3 and Pol-delta4 process Okazaki fragments in vitro, Pol-delta3 may also be better suited to fulfill this task, exhibiting near-absence of strand displacement activity compared to Pol-delta4 and stalling on encounter with the 5'-blocking oligonucleotides. Pol-delta3 idling process may avoid the formation of a gap, while maintaining a nick that can be readily ligated. Along with DNA polymerase kappa, DNA polymerase delta carries out approximately half of nucleotide excision repair (NER) synthesis following UV irradiation. In this context, POLD3, along with PCNA and RFC1-replication factor C complex, is required to recruit POLD1, the catalytic subunit of the polymerase delta complex, to DNA damage sites. Under conditions of DNA replication stress, required for the repair of broken replication forks through break-induced replication (BIR). Involved in the translesion synthesis (TLS) of templates carrying O6-methylguanine or abasic sites performed by Pol-delta4, independently of DNA polymerase zeta (REV3L) or eta (POLH). Facilitates abasic site bypass by DNA polymerase delta by promoting extension from the nucleotide inserted opposite the lesion. Also involved in TLS, as a component of the tetrametric DNA polymerase zeta complex. Along with POLD2, dramatically increases the efficiency and processivity of DNA synthesis of the DNA polymerase zeta complex compared to the minimal zeta complex, consisting of only REV3L and REV7.
Protein Name Dna Polymerase Delta Subunit 3
Dna Polymerase Delta Subunit C
Dna Polymerase Delta Subunit P66
Dna Polymerase Delta Subunit P68
Database Links Reactome: R-HSA-110314
Reactome: R-HSA-174411
Reactome: R-HSA-174414
Reactome: R-HSA-174417
Reactome: R-HSA-174437
Reactome: R-HSA-5358565
Reactome: R-HSA-5358606
Reactome: R-HSA-5651801
Reactome: R-HSA-5656169
Reactome: R-HSA-5685942
Reactome: R-HSA-5696397
Reactome: R-HSA-5696400
Reactome: R-HSA-6782135
Reactome: R-HSA-6782210
Reactome: R-HSA-69091
Reactome: R-HSA-69166
Reactome: R-HSA-69183
Cellular Localisation Cytoplasm
Nucleus
Partially Colocalizes With Pcna And Pold1 At S Phase Replication Sites
Recruited To Dna Damage Sites Within 2 Hours Following Uv Irradiation
Alternative Antibody Names Anti-Dna Polymerase Delta Subunit 3 antibody
Anti-Dna Polymerase Delta Subunit C antibody
Anti-Dna Polymerase Delta Subunit P66 antibody
Anti-Dna Polymerase Delta Subunit P68 antibody
Anti-POLD3 antibody
Anti-KIAA0039 antibody

Information sourced from Uniprot.org

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