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MPG / AAG Recombinant Rabbit Monoclonal Antibody [PSH06-34]

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Specification

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Catalog# HA722668

MPG / AAG Recombinant Rabbit Monoclonal Antibody [PSH06-34]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

MPG / AAG Recombinant Rabbit Monoclonal Antibody [PSH06-34]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human MPG / AAG aa 1-298.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell, FC

Target Molecular Weight

Predicted band size: 33 kDa

Positive Control

HeLa cell lysate, HEK-293 cell lysate, Jurkat cell lysate, MCF7 cell lysate, K-562 cell lysate, U-937 cell lysate, LoVo cell lysate, HL-60 cell lysate, Raji cell lysate, HepG2 cell lysate, MCF7-si NT cell lysate, MCF7-si 商品名 cell lysate, human breast tissue, human colon tissue, mouse breast tissue, mouse colon tissue, rat breast tissue, rat colon tissue, K-562.

Conjugation

unconjugated

Clone Number

PSH06-34

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

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

DNA-3-methyladenine glycosylase also known as 3-alkyladenine DNA glycosylase (AAG) or N-methylpurine DNA glycosylase (MPG) is an enzyme that in humans is encoded by the MPG gene. Alkyladenine DNA glycosylase is a specific type of DNA glycosylase. This subfamily of monofunctional glycosylases is involved in the recognition of a variety of base lesions, including alkylated and deaminated purines, and initiating their repair via the base excision repair pathway. To date, the human AAG (hAAG) is the only glycosylase identified that excises alkylation-damaged purine bases in human cells.

Background References

1. Pinthong N et al. Molecular characterization of Plasmodium falciparum DNA-3-methyladenine glycosylase. Malar J. 2020 Aug

2. de Barrios O et al. ZEB1 promotes inflammation and progression towards inflammation-driven carcinoma through repression of the DNA repair glycosylase MPG in epithelial cells. Gut. 2019 Dec

Subcellular Location

Cytoplasm, Mitochondrion matrix, mitochondrion nucleoid, Nucleus.

UNIPROT

SwissProt: P29372 Human

SwissProt: Q04841 Mouse

SwissProt: P23571 Rat

Synonyms

3 alkyladenine DNA glycosylase antibody

3-alkyladenine DNA glycosylase antibody

3-methyladenine DNA glycosidase antibody

3MG_HUMAN antibody

AAG antibody

ADPG antibody

Alkyladenine DNA glycosylase antibody

anpg antibody

APNG antibody

CRA36.1 antibody

Expand

Images

  • Western blot analysis of MPG / AAG on different lysates with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HEK-293 cell lysate<br />Lane 3: Jurkat cell lysate<br />Lane 4: MCF7 cell lysate<br />Lane 5: K-562 cell lysate<br />Lane 6: U-937 cell lysate<br />Lane 7: LoVo cell lysate<br />Lane 8: HL-60 cell lysate<br />Lane 9: Raji cell lysate<br />Lane 10: HepG2 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 33 kDa<br />Observed band size: 35 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of MPG / AAG on different lysates with Rabbit anti-MPG / AAG antibody (HA722668) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HEK-293 cell lysate
    Lane 3: Jurkat cell lysate
    Lane 4: MCF7 cell lysate
    Lane 5: K-562 cell lysate
    Lane 6: U-937 cell lysate
    Lane 7: LoVo cell lysate
    Lane 8: HL-60 cell lysate
    Lane 9: Raji cell lysate
    Lane 10: HepG2 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 33 kDa
    Observed band size: 35 kDa

    Exposure time: 20 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722668) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution.<br /><br />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 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution for 1 hour 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 tissue with Rabbit anti-MPG / AAG antibody (HA722668) at 1/1,000 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722668) at 1/1,000 dilution for 1 hour 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 colon tissue with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution.<br /><br />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 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution for 1 hour 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 colon tissue with Rabbit anti-MPG / AAG antibody (HA722668) at 1/1,000 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722668) at 1/1,000 dilution for 1 hour 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 breast tissue with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution.<br /><br />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 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution for 1 hour 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 breast tissue with Rabbit anti-MPG / AAG antibody (HA722668) at 1/1,000 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722668) at 1/1,000 dilution for 1 hour 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 colon tissue with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution.<br /><br />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 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution for 1 hour 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 colon tissue with Rabbit anti-MPG / AAG antibody (HA722668) at 1/1,000 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722668) at 1/1,000 dilution for 1 hour 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 breast tissue with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution.<br /><br />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 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution for 1 hour 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 breast tissue with Rabbit anti-MPG / AAG antibody (HA722668) at 1/1,000 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722668) at 1/1,000 dilution for 1 hour 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 colon tissue with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution.<br /><br />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 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution for 1 hour 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 colon tissue with Rabbit anti-MPG / AAG antibody (HA722668) at 1/1,000 dilution.

    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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722668) at 1/1,000 dilution for 1 hour 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.

  • Immunocytochemistry analysis of K-562 cells labeling MPG / AAG with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/100 dilution in 1% BSA in PBST 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. PBS instead of the primary antibody was used as the secondary antibody only control. 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 1/100 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 MPG / AAG with Rabbit anti-MPG / AAG antibody (HA722668) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MPG / AAG antibody (HA722668) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of K-562 cells labeling MPG / AAG.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of K-562 cells labeling MPG / AAG.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722668, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of MPG / AAG on different lysates with Rabbit anti-MPG / AAG antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution.<br /><br />Lane 1: MCF7-si NT cell lysate<br />Lane 2: MCF7-si MPG / AAG cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 33 kDa<br />Observed band size: 35 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722668" style="font-weight: bold;text-decoration: underline;">HA722668</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of MPG / AAG on different lysates with Rabbit anti-MPG / AAG antibody (HA722668) at 1/1,000 dilution.

    Lane 1: MCF7-si NT cell lysate
    Lane 2: MCF7-si MPG / AAG cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 33 kDa
    Observed band size: 35 kDa

    Exposure time: 10 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722668) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"