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Knockout Tested Rabbit Recombinant Monoclonal POLE antibody. Suitable for WB, IP, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

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Images

Immunoprecipitation - Anti-POLE antibody [EPR28639-64] (AB318968), expandable thumbnail
  • Immunoprecipitation - Anti-POLE antibody [EPR28639-64] (AB318968), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-POLE antibody [EPR28639-64] (AB318968), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-POLE antibody [EPR28639-64] (AB318968), expandable thumbnail
  • Western blot - Anti-POLE antibody [EPR28639-64] (AB318968), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIPICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Not recommended
Mouse
Tested
Tested
Expected
Expected
Not recommended
Rat
Tested
Expected
Expected
Expected
Not recommended

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Species
Mouse
Dilution info
1/30
Notes

-

Expected
Expected

Species
Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human
Dilution info
1/500
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human
Dilution info
1/50
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Target data

Function

Catalytic component of the DNA polymerase epsilon complex (PubMed:10801849). Participates in chromosomal DNA replication (By similarity). Required during synthesis of the leading DNA strands at the replication fork, binds at/or near replication origins and moves along DNA with the replication fork (By similarity). Has 3'-5' proofreading exonuclease activity that corrects errors arising during DNA replication (By similarity). Involved in DNA synthesis during DNA repair (PubMed:20227374, PubMed:27573199). Along with DNA polymerase POLD1 and DNA polymerase POLK, has a role in excision repair (NER) synthesis following UV irradiation (PubMed:20227374).

Alternative names

POLE1, POLE, DNA polymerase epsilon catalytic subunit A, 3'-5' exodeoxyribonuclease, DNA polymerase II subunit A

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal POLE antibody. Suitable for WB, IP, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR28639-64
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The POLE protein also known as DNA polymerase epsilon catalytic subunit A functions as an important component in DNA replication and repair mechanisms. POLE possesses a molecular mass of approximately 261 kDa and demonstrates expression primarily in proliferating cells such as those in the bone marrow lymphoid tissues and the gastrointestinal tract. It operates within the eukaryotic DNA polymerase family distinct from other polymerases due to its high fidelity in DNA synthesis which minimizes errors during replication.

Biological function summary

The POLE protein plays an essential role in the high-fidelity synthesis of leading-strand DNA during replication. POLE is part of a larger complex known as the DNA polymerase epsilon holoenzyme collaborating with accessory proteins that enhance its ability to synthesize DNA accurately. Moreover this protein contributes significantly to the proofreading function as it has intrinsic 3' to 5' exonuclease activity allowing the removal of incorrectly paired nucleotides therefore preventing mutations.

Pathways

The POLE protein is fundamentally involved in the DNA replication and repair pathways. These pathways ensure genomic integrity and stability important for normal cell division and preventing genomic instability associated with cancer. POLE interacts with other proteins such as PCNA and RFC (replication factor C) within these pathways facilitating efficient replication fork progression and repair processes. POLE's activity ensures coordination with leading strand synthesis complementing the functions of other polymerases involved in lagging strand DNA synthesis.

Associated diseases and disorders

Mutations in the POLE gene have been linked to colorectal cancer and endometrial cancer. These conditions are often characterized by a hypermutated phenotype arising from impaired DNA proofreading ability due to ineffective exonuclease activity. In this context POLE mutations can assume significance in sporadic cancer cases as well as in hereditary cancer syndromes where it interacts with mismatch repair proteins highlighting its role in maintaining genomic fidelity.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Immunoprecipitation - Anti-POLE antibody [EPR28639-64] (ab318968), expandable thumbnail

    Immunoprecipitation - Anti-POLE antibody [EPR28639-64] (ab318968)

    POLE was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab318968 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318968 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate

    Lane 2: ab318968 IP in K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab318968 in K-562 whole cell lysate

    The bands beneath the target band between 100 kDa and 260 kDa are likely to be degraded target fragments.

    All lanes: Immunoprecipitation - Anti-POLE antibody [EPR28639-64] (ab318968) at 1/30 dilution

    Lanes 1 - 2: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 10 µg with NFDM/TBST

    Lanes 4 - 5: F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 10 µg with NFDM/TBST

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Exposure time: 10s

  • Immunoprecipitation - Anti-POLE antibody [EPR28639-64] (ab318968), expandable thumbnail

    Immunoprecipitation - Anti-POLE antibody [EPR28639-64] (ab318968)

    POLE was immunoprecipitated from 0.35 mg parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate with ab318968 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318968 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate

    Lane 2: ab318968 IP in parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab318968 in parental HAP1(HOWT01) whole cell lysate

    The bands beneath the target band between 100 kDa and 260 kDa are likely to be degraded target fragments.

    All lanes: Immunoprecipitation - Anti-POLE antibody [EPR28639-64] (ab318968) at 1/30 dilution

    Lanes 1 - 2: parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 10 µg with NFDM/TBST

    Lanes 4 - 5: POLE Knockout HAP1(HO010404) whole cell lysate at 10 µg with NFDM/TBST

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Exposure time: 6s

  • Flow Cytometry (Intracellular) - Anti-POLE antibody [EPR28639-64] (ab318968), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-POLE antibody [EPR28639-64] (ab318968)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast) cells labelling POLE with ab318968 at 1/50 dilution (1 ug)/Magenta compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

  • Flow Cytometry (Intracellular) - Anti-POLE antibody [EPR28639-64] (ab318968), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-POLE antibody [EPR28639-64] (ab318968)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Parental HAP1(human chronic myelogenous leukemia near-haploid cell, Left) / POLE KO HAP1(Right) cells labelling POLE with ab318968 at 1/50 dilution (1ug) / Magenta compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

  • Western blot - Anti-POLE antibody [EPR28639-64] (ab318968), expandable thumbnail

    Western blot - Anti-POLE antibody [EPR28639-64] (ab318968)

    The identity of the band at 100 kDa is unknown.

    The bands beneath the target band between 100 kDa and 260 kDa (Lane 1-8) are likely to be degraded target fragments.

    In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] ab129002) staining at 1/10000 dilution.

    Exposure time: Lane 1-8: 15 seconds; Lane 9-12: 48 seconds

    All lanes: Western blot - Anti-POLE antibody [EPR28639-64] (ab318968) at 1/1000 dilution

    Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

    Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

    Lane 3: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

    Lane 4: THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg with NFDM/TBST

    Lane 5: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg with NFDM/TBST

    Lane 6: F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

    Lane 7: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg with NFDM/TBST

    Lane 8: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg with NFDM/TBST

    Lane 9: HeLa (human cervical adenocarcinoma epithelial cell) whole cell fresh lysate at 20 µg with NFDM/TBST

    Lane 10: 293T (human embryonic kidney epithelial cell) whole cell fresh lysate at 20 µg with NFDM/TBST

    Lane 11: NIH/3T3 (mouse embryonic fibroblast) whole cell fresh lysate at 20 µg with NFDM/TBST

    Lane 12: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell fresh lysate at 20 µg with NFDM/TBST

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 260 kDa, 124 kDa

  • Western blot - Anti-POLE antibody [EPR28639-64] (ab318968), expandable thumbnail

    Western blot - Anti-POLE antibody [EPR28639-64] (ab318968)

    The identity of the band at 100 kDa is unknown.

    The bands beneath the target band between 100 kDa and 260 kDa (Lane 1-4) are likely to be degraded target fragments.

    In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] ab129002) staining at 1/10000 dilution.

    Exposure time: Lane 1-4: 15 seconds; Lane 4: 180 seconds

    All lanes: Western blot - Anti-POLE antibody [EPR28639-64] (ab318968) at 1/1000 dilution

    Lane 1: Mouse spleen tissue lysate at 20 µg with NFDM/TBST

    Lane 2: Mouse testis tissue lysate at 20 µg with NFDM/TBST

    Lane 3: Mouse placenta tissue lysate at 20 µg with NFDM/TBST

    Lane 4: Rat testis tissue lysate at 20 µg with NFDM/TBST

    Lane 5: Mouse testis fresh tissue lysate at 20 µg with NFDM/TBST

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 260 kDa, 124 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-POLE antibody [EPR28639-64] (ab318968), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-POLE antibody [EPR28639-64] (ab318968)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized POLE KO HAP 1 (POLE knockout human chronic myelogenous leukemia near-haploid cell) cells labelling POLE with ab318968 at 1/500 (1.07 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

    Confocal image showing nuclear staining in wildtype HAP1 cells and negative staining in POLE knockout HAP1 cells(shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

  • Western blot - Anti-POLE antibody [EPR28639-64] (ab318968), expandable thumbnail

    Western blot - Anti-POLE antibody [EPR28639-64] (ab318968)

    The identity of the band at 100 kDa is unknown.

    The bands beneath the target band between 100 kDa and 260 kDa are likely to be degraded target fragments.

    In Western blot, ab318968 was shown to bind specifically to POLE. Target of interest was observed at 260kDa in wild-type HAP1 cell lysates (Lane 1) with no signal observed at this size in POLE knockout cell line (Lane 2).

    In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] ab129002) staining at 1/10000 dilution.

    All lanes: Western blot - Anti-POLE antibody [EPR28639-64] (ab318968) at 1/1000 dilution

    Lane 1: parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 20 µg with NFDM/TBST

    Lane 2: POLE Knockout HAP1(HO010404) whole cell lysate at 20 µg with NFDM/TBST

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 260 kDa, 124 kDa

    Exposure time: 180s

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Product protocols

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