POLD1 Antibody (YA1718)(PBS only)
(Synonyms: CDC2; CRCS10; MDPL; POLD; POLD1)POLD1 Antibody (YA1718) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to POLD1.
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Host:
Rabbit
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Isotype:
IgG
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Application:
WB, IHC-P, ICC/IF, FC
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Reactivity :
Human, Mouse, Rat
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Formulation:
Supplied in PBS, pH 7.4.
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Conjugation:
Non-conjugated
Applications
| Application |
WB
WB: Western Blot
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ICC/IF
ICC/IF: Immunocytochemistry/
Immunofluorescence |
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| Dilution Ratio | 1:500-1:1000 | 1:50-1:200 |
Product Details
POLD1 Antibody (YA1718) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to POLD1.
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Host Rabbit
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Species ReactivityHuman, Mouse, Rat
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Observed Molecular WeightObserved band size: 124 kDaNote: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
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Calculated Molecular Weight Predicted band size: 124 kDa
A synthetic peptide of human DNA Polymerase delta, catalytic subunit
Affinity Purified
Non-conjugated
IgG
Product Properties
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Appearance
Liquid
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Formulation
Supplied in PBS, pH 7.4.
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Storage & Stability
Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
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Shipping
Shipping with blue ice.
Verification Images
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Western blot analysis of extracts from Hela (lane 2, 20ug), MCF-7 (lane 3, 20ug), HEK293T (lane 4, 20ug) and RAW264.7 (lane 5, 20ug) using POLD1 Antibody (HY-P81973). Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST at 4°C overnight. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80993, 1/10,000) was used in 5% non-fat milk in TBST for 2 hour at room temperature. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HY-P8001, 1/10,000) was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue using POLD1 Antibody (YA1718). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81973, 1/100) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam.
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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using POLD1 Antibody (YA1718). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81973, 1/100) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam.
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Immunohistochemical analysis of paraffin-embedded human liver cancer tissue using POLD1 Antibody (YA1718). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81973, 1/100) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam.
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Immunocytochemistry analysis of Hela cells labeling POLD1 with POLD1 Antibody (HY-P81973) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with POLD1 Antibody (HY-P81973) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
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Immunocytochemistry analysis of U2OS cells labeling POLD1 with POLD1 Antibody (HY-P81973) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with POLD1 Antibody (HY-P81973) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
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Tyramide signaling amplification based immunofluorescence analysis of paraffin-embedded human ovarian cancer tissue using POLD1 Antibody (YA1718). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81973, 1/300) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer and fluorescent tyramide signal amplification system.Immunostaining was performed using a Bicolor signal amplification fluorescence staining kit. Tissues were counterstained with DAPI (blue) and mounted with Anti-fade fluorescence mounting medium.
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Tyramide signaling amplification based immunofluorescence analysis of paraffin-embedded human cervical cancer tissue using POLD1 Antibody (YA1718). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81973, 1/400) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer and fluorescent tyramide signal amplification system.Immunostaining was performed with Vari Fluor 594 TSA (200×)(HY-D1835). Tissues were counterstained with DAPI (blue) and mounted with Anti-fade fluorescence mounting medium.
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Tyramide signaling amplification based immunofluorescence analysis of paraffin-embedded human colon cancer tissue using POLD1 Antibody (YA1718). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81973, 1/400) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer and fluorescent tyramide signal amplification system.Immunostaining was performed with Vari Fluor 594 TSA (200×)(HY-D1835). Tissues were counterstained with DAPI (blue) and mounted with Anti-fade fluorescence mounting medium.
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Tyramide signaling amplification based immunofluorescence analysis of paraffin-embedded human liver cancer tissue using POLD1 Antibody (YA1718). The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P81973, 1/400) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer and fluorescent tyramide signal amplification system.Immunostaining was performed with Vari Fluor 594 TSA (200×)(HY-D1835). Tissues were counterstained with DAPI (blue) and mounted with Anti-fade fluorescence mounting medium.
Background
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Function
POLD1 is an As the catalytic component of the trimeric (Pol-delta3 complex) and tetrameric DNA polymerase delta complexes (Pol-delta4 complex), plays a crucial role in high fidelity genome replication, including in lagging strand synthesis, and repair. Exhibits both DNA polymerase and 3'- to 5'-exonuclease activities. Requires the presence of accessory proteins POLD2, POLD3 and POLD4 for full activity. Depending upon the absence (Pol-delta3) or the presence of POLD4 (Pol-delta4), displays differences in catalytic activity. Most notably, expresses higher proofreading activity in the context of Pol-delta3 compared with that of Pol-delta4. Although both Pol-delta3 and Pol-delta4 process Okazaki fragments in vitro, Pol-delta3 may 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. Under conditions of DNA replication stress, in the presence of POLD3 and POLD4, may catalyze the repair of broken replication forks through break-induced replication (BIR). Involved in the translesion synthesis (TLS) of templates carrying O6-methylguanine, 8oxoG or abasic sites[1][2][3][4][5][6][7][8][9].
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Subcellular Localization
Nucleus
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Expression
Tissue_specificity:Widely expressed, with high levels of expression in heart and lung
Induction:Up-regulated by serum stimulation -
Subunit
Component of the tetrameric DNA polymerase delta complex (Pol-delta4), which consists of POLD1/p125, POLD2/p50, POLD3/p66/p68 and POLD4/p12, with POLD1 bearing both DNA polymerase and 3' to 5' proofreading exonuclease activities (PubMed:11595739, PubMed:12522211, PubMed:17317665, PubMed:22801543, PubMed:31449058, PubMed:31629014). Within Pol-delta4, directly interacts with POLD2 and POLD4 (PubMed:11328591, PubMed:12403614, PubMed:16510448). Following genotoxic stress by DNA-damaging agents, such as ultraviolet light and methyl methanesulfonate, or by replication stress induced by treatment with hydroxyurea or aphidicolin, Pol-delta4 is converted into a trimeric form of the complex (Pol-delta3) by POLD4 degradation. Pol-delta3 is the major form at S phase replication sites and DNA damage sites (PubMed:17317665, PubMed:22801543). POLD1 displays different catalytic properties depending upon the complex it is found in (PubMed:17317665). It exhibits higher proofreading activity and fidelity than Pol-delta4, making it particularly well suited to respond to DNA damage (PubMed:19074196, PubMed:20334433). Directly interacts with PCNA, as do POLD3 and POLD4; this interaction stimulates Pol-delta4 polymerase activity (PubMed:11328591, PubMed:12403614, PubMed:12522211, PubMed:16510448, PubMed:24022480, PubMed:24939902). As POLD2 and POLD4, directly interacts with WRNIP1; this interaction stimulates DNA polymerase delta-mediated DNA synthesis, independently of the presence of PCNA. This stimulation may be due predominantly to an increase of initiation frequency and also to increased processivity (PubMed:15670210). Also observed as a dimeric complex with POLD2 (Pol-delta2 complex). Pol-delta2 is relatively insensitive to the PCNA stimulation (2-5-fold) compared to Pol-delta4 that is stimulated by over 50-fold (PubMed:12403614). The DNA polymerase delta complex interacts with POLDIP2; this interaction is probably mediated through direct binding to POLD2 (PubMed:12522211). Interacts with CIAO1 (PubMed:23891004). Interacts with POLDIP2 (PubMed:24191025). Interacts with RFC1 (PubMed:31629014)
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SwissProt ID
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Synonyms
CDC2; CRCS10; MDPL; POLD; POLD1
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Research Field
Epigenetics and Nuclear Signaling
Documentation