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DNA Polymerase beta Recombinant Rabbit Monoclonal Antibody [JM93-12]

Usd: 385

Specification

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Catalog# ET1705-12

DNA Polymerase beta Recombinant Rabbit Monoclonal Antibody [JM93-12]

Application

  • WB

  • IHC-P

  • IF-Cell

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

  • Zebrafish

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

DNA Polymerase beta Recombinant Rabbit Monoclonal Antibody [JM93-12]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human DNA Polymerase beta aa 286-335 / 335.

Species Reactivity

Human, Mouse, Rat, Zebrafish

Validated Applications

WB, IHC-P, IF-Cell, IP

Target Molecular Weight

Predicted band size: 38 kDa

Positive Control

HeLa (Human cervical adenocarcinoma cell) cell lysate, SH-SY5Y (Human neuroblastoma cell) cell lysate, HT-29 (Human colorectal adenocarcinoma cell) cell lysate, Neuro-2a (Mouse brain neuroblastoma cell) cell lysate, C6 (Rat glioma cell) cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, K-562, rat lung tissue, human breast carcinoma tissue, human stomach carcinoma tissue, human uterus tissue, mouse cerebellum tissue.

Conjugation

unconjugated

Clone Number

JM93-12

RRID

AB_3070544

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:50-1:200

  • IF-Cell

  • 1:20-1:50

  • IP

  • 1:10-1:20

Target

Function

DNA polymerase beta, also known as POLB, is an enzyme present in eukaryotes. In humans, it is encoded by the POLB gene. In eukaryotic cells, DNA polymerase beta (POLB) performs base excision repair (BER) required for DNA maintenance, replication, recombination, and drug resistance. The mitochondrial DNA of mammalian cells is constantly under attack from oxygen radicals released during ATP production. Mammalian cell mitochondria contain an efficient base excision repair system employing POLB that removes some frequent oxidative DNA damages. POLB thus has a key role in maintaining the stability of the mitochondrial genome. An analysis of the fidelity of DNA replication by polymerase beta in the neurons from young and very aged mice indicated that aging has no significant effect on the fidelity of DNA synthesis by polymerase beta. This finding was considered to provide evidence against the error catastrophe theory of aging.

Background References

1. Howard MJ. et. al. DNA polymerase beta nucleotide-stabilized template misalignment fidelity depends on local sequence context. J Biol Chem. 2020 Jan

2. Kurosawa A. et. al. Complex genetic interactions between DNA polymerase beta and the NHEJ ligase. FEBS J. 2020 Jan

Sequence Similarity

Belongs to the DNA polymerase type-X family.

Post-translational Modification

Methylation by PRMT6 stimulates the polymerase activity by enhancing DNA binding and processivity.; Ubiquitinated at Lys-41, Lys-61 and Lys-81: monoubiquitinated by HUWE1/ARF-BP1. Monoubiquitinated protein is then the target of STUB1/CHIP, which catalyzes polyubiquitination from monoubiquitin, leading to degradation by the proteasome. USP47 mediates the deubiquitination of monoubiquitinated protein, preventing polyubiquitination by STUB1/CHIP and its subsequent degradation.

Subcellular Location

Nucleus, Cytoplasm.

UNIPROT

SwissProt: P06746 Human

SwissProt: Q8K409 Mouse

SwissProt: P06766 Rat

Synonyms

DNA directed DNA polymerase beta antibody

DNA pol beta antibody

DNA polymerase beta antibody

DNA polymerase beta subunit antibody

DPOLB_HUMAN antibody

MGC125976 antibody

Pol B antibody

Pol beta antibody

POLB antibody

Polymerase (DNA directed) beta antibody

Images

  • Western blot analysis of DNA Polymerase beta on different lysates with Rabbit anti-DNA Polymerase beta antibody (<a href="/products/ET1705-12" style="font-weight: bold;text-decoration: underline;">ET1705-12</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate<br />Lane 2: SH-SY5Y (Human neuroblastoma cell) cell lysate<br />Lane 3: HT-29 (Human colorectal adenocarcinoma cell) cell lysate<br />Lane 4: Neuro-2a (Mouse brain neuroblastoma cell) cell lysate<br />Lane 5: C6 (Rat glioma cell) cell lysate<br />Lane 6: Mouse brain tissue lysate<br />Lane 7: Rat brain tissue lysate<br /><br />Cell Lysates at 20 µg/Lane, Tissue Lysates at 40 µg/Lane.<br />Exposure time: 25 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1705-12" style="font-weight: bold;text-decoration: underline;">ET1705-12</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 38 kDa<br />Observed band size: 38 kDa

    Western blot analysis of DNA Polymerase beta on different lysates with Rabbit anti-DNA Polymerase beta antibody (ET1705-12) at 1/5,000 dilution.

    Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
    Lane 2: SH-SY5Y (Human neuroblastoma cell) cell lysate
    Lane 3: HT-29 (Human colorectal adenocarcinoma cell) cell lysate
    Lane 4: Neuro-2a (Mouse brain neuroblastoma cell) cell lysate
    Lane 5: C6 (Rat glioma cell) cell lysate
    Lane 6: Mouse brain tissue lysate
    Lane 7: Rat brain tissue lysate

    Cell Lysates at 20 µg/Lane, Tissue Lysates at 40 µg/Lane.
    Exposure time: 25 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1705-12, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 38 kDa
    Observed band size: 38 kDa

  • Immunocytochemistry analysis of K-562 cells labeling DNA Polymerase beta with Rabbit anti-DNA Polymerase beta antibody (<a href="/products/ET1705-12" style="font-weight: bold;text-decoration: underline;">ET1705-12</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-DNA Polymerase beta antibody (<a href="/products/ET1705-12" style="font-weight: bold;text-decoration: underline;">ET1705-12</a>) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of K-562 cells labeling DNA Polymerase beta with Rabbit anti-DNA Polymerase beta antibody (ET1705-12) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-DNA Polymerase beta antibody (ET1705-12) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat lung tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-12" style="font-weight: bold;text-decoration: underline;">ET1705-12</a>, 1/50) for 30 minutes at room temperature. 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 DPX.

    Immunohistochemical analysis of paraffin-embedded rat lung tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-12, 1/50) for 30 minutes at room temperature. 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 DPX.

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-12" style="font-weight: bold;text-decoration: underline;">ET1705-12</a>, 1/50) for 30 minutes at room temperature. 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 DPX.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-12, 1/50) for 30 minutes at room temperature. 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 DPX.

  • Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-12" style="font-weight: bold;text-decoration: underline;">ET1705-12</a>, 1/50) for 30 minutes at room temperature. 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 DPX.

    Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-12, 1/50) for 30 minutes at room temperature. 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 DPX.

  • Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-12" style="font-weight: bold;text-decoration: underline;">ET1705-12</a>, 1/50) for 30 minutes at room temperature. 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 DPX.

    Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-12, 1/50) for 30 minutes at room temperature. 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 DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-12" style="font-weight: bold;text-decoration: underline;">ET1705-12</a>, 1/50) for 30 minutes at room temperature. 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 DPX.

    Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue using anti-DNA Polymerase beta antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-12, 1/50) for 30 minutes at room temperature. 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 DPX.

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