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3mC Recombinant Rabbit Monoclonal Antibody [PSH03-21] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750861

3mC Recombinant Rabbit Monoclonal Antibody [PSH03-21] - BSA and Azide free

Application

  • IF-Cell

  • Dot Blot

Reactivity

  • Species independent

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

3mC Recombinant Rabbit Monoclonal Antibody [PSH03-21] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

3-mc-OVA

Species Reactivity

Species independent

Validated Applications

IF-Cell, Dot Blot

Positive Control

U-2 OS cells treated with 4mM Methyl Methanesulfonate for 1 hour.

Conjugation

unconjugated

Clone Number

PSH03-21

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • IF-Cell

  • 1:100

  • Dot Blot

  • 1:2,000

Target

Function

Methylation of DNA can occur non-enzymatically at the nitrogen-three of the cytosine base through spontaneous exposure to endogenous S-adenosyl methionine (SAM). The resulting 3-methylcytosine (3-mC) is mutagenic and must be repaired, which occurs in humans through the base excision repair (BER) or dealkylation via human homologues of the E. coli AlkB protein. 3-methylcytosine is present in human cell lines and increased levels of 3-mC impair proliferation.

Background References

暂无

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of U-2 OS cells treated with 4mM Methyl Methanesulfonate for 1 hour labeling 3mC with Rabbit anti-3mC antibody (<a href="/products/HA750861" style="font-weight: bold;text-decoration: underline;">HA750861</a>) at 1/100 dilution.<br /><br />Cells were fixed in 70% ethyl alcohol for 5 minutes at room temperature, then subjected to acid hydrolysis using 2M HCl in TBST for 30 minutes at room temperature. Permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-3mC antibody (<a href="/products/HA750861" style="font-weight: bold;text-decoration: underline;">HA750861</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.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of U-2 OS cells treated with 4mM Methyl Methanesulfonate for 1 hour labeling 3mC with Rabbit anti-3mC antibody (HA750861) at 1/100 dilution.

    Cells were fixed in 70% ethyl alcohol for 5 minutes at room temperature, then subjected to acid hydrolysis using 2M HCl in TBST for 30 minutes at room temperature. Permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-3mC antibody (HA750861) 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.

  • Dot blot analysis of 3mC on different proteins with Rabbit anti-3mC antibody (<a href="/products/HA750861" style="font-weight: bold;text-decoration: underline;">HA750861</a>) at 1/2,000 dilution. 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 for 1 hour at room temperature.<br /><br />Lane 1: Uridine-BSA (negative)<br />Lane 2: Cytidine-BSA (negative)<br />Lane 3: 5-Methylcytosine-BSA (negative)<br />Lane 4: 3-Methylcytosine-BSA (positive)<br />Lane 5: 5-Hydroxymethylcytidine-BSA (negative)<br /><br />Proteins loading: 50ng, 25ng, 12.5ng, 6.25ng;<br /><br />Blocking and dilution buffer: 5% NFDM/TBST;<br /><br />Exposure time: 1 minute.

    Dot blot analysis of 3mC on different proteins with Rabbit anti-3mC antibody (HA750861) at 1/2,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature.

    Lane 1: Uridine-BSA (negative)
    Lane 2: Cytidine-BSA (negative)
    Lane 3: 5-Methylcytosine-BSA (negative)
    Lane 4: 3-Methylcytosine-BSA (positive)
    Lane 5: 5-Hydroxymethylcytidine-BSA (negative)

    Proteins loading: 50ng, 25ng, 12.5ng, 6.25ng;

    Blocking and dilution buffer: 5% NFDM/TBST;

    Exposure time: 1 minute.

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