AB108989

重组Anti-MGMT抗体[EPR4398]

Anti-MGMT antibody [EPR4398]

20ul selling size
RabMAb
Recombinant
KO Validated
了解详情

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(1 Publication)

Rabbit Recombinant Monoclonal MGMT antibody. Suitable for IP, WB and reacts with Human samples. Cited in 1 publication.

查看别名

Methylated-DNA--protein-cysteine methyltransferase, 6-O-methylguanine-DNA methyltransferase, O-6-methylguanine-DNA-alkyltransferase, MGMT

5 Images

Unknown

Immunoprecipitation - Anti-MGMT antibody [EPR4398] (AB108989)

ab108989 (purified) at 1/30 immunoprecipitating MGMT in 10 μg MCF7 (Lanes 1 and 2, observed at 23 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-MGMT antibody [EPR4398] (ab108989)

Predicted band size: 22 kDa

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Western blot - Anti-MGMT antibody [EPR4398] (AB108989)

All lanes:

Western blot - Anti-MGMT antibody [EPR4398] (ab108989) at 1/1000 dilution

Lane 1:

MCF7 cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 22 kDa

Observed band size: 23 kDa

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Lab

Western blot - Anti-MGMT antibody [EPR4398] (AB108989)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-MGMT antibody [EPR4398] (ab108989) at 1/1000 dilution

Lane 1:

MCF7 cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 23 kDa

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Lab

Western blot - Anti-MGMT antibody [EPR4398] (AB108989)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : MGMT knockout HAP1 cell lysate (20 μg)
Lane 3 : MCF7 cell lysate (20 μg)
Lane 4 : Jurkat cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab108989 observed at 22 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab108989 was shown to specifically react with MGMT when MGMT knockout samples were used. Wild-type and MGMT knockout samples were subjected to SDS-PAGE. ab108989 and ab8245 (loading control to MGMT) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-MGMT antibody [EPR4398] (ab108989)

Predicted band size: 22 kDa

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Lab

Western blot - Anti-MGMT antibody [EPR4398] (AB108989)

Western blot : Anti-MGMT antibody [EPR4398] (ab108989) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab108989 was shown to bind specifically to MGMT. A band was observed at 22 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in MGMT CRISPR-Cas9 edited cell line ab286541. The band observed in the CRISPR-Cas9 edited lysate lane below 22 kDa is likely to represent a truncated form of MGMT. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MGMT CRISPR-Cas9 edited HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-MGMT antibody [EPR4398] (ab108989) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human MGMT knockout HCT116 cell line (<a href='/products/cell-lines/human-mgmt-knockout-hct116-cell-line-ab286541'>ab286541</a>)

Lane 2:

MGMT CRISPR-Cas9 edited HCT 116 cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 22 kDa

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不同偶联物与剂型 (1)

Carrier free

Anti-MGMT antibody [EPR4398] - BSA and Azide free

关键信息

宿主种属

Rabbit

克隆

Monoclonal

克隆号

EPR4398

亚型

IgG

不含载体蛋白

反应种属

Human

应用

WB, IP

applications

免疫原

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

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反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/10 - 1/100", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000 - 1/10000", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Mouse": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Rat": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }

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产品详情

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

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

What are the advantages of a recombinant monoclonal antibody?
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.

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性能和储存信息

形式

Liquid

纯化工艺

Affinity purification Protein A

存储溶液

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

运输条件

Blue Ice

分装信息

Upon delivery aliquot

储存信息

Stable for 12 months at -20°C

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补充信息

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

MGMT also known as O-6-methylguanine-DNA methyltransferase is an enzyme with a significant role in DNA repair. MGMT protein has a molecular weight of about 22 kDa. This protein is typically expressed in various tissues including the liver kidney and brain where it functions to protect cells from mutagenic and cytotoxic alterations. MGMT works by transferring alkyl groups from the O6 position of guanine to a cysteine residue within its active site. This mechanism directly reverses DNA damage maintaining genomic stability.

Biological function summary

MGMT prevents mutations by reversing alkylation damage playing a significant role in preserving DNA integrity. It acts independently and does not form part of any larger protein complexes. This enzyme-mediated repair is central to cellular defense against mutagenic threats such as alkylating agents. By curbing DNA damage MGMT can thwart the onset of harmful genetic mutations that might lead to issues like cancer development.

Pathways

MGMT is intimately involved in DNA damage response and repair pathways. Notably it interacts with base excision repair and direct reversal repair pathways. MGMT's function is distinct but complementary to the activities of other DNA repair proteins like XRCC1 and APE1 which are involved in the base excision repair pathway. The presence of MGMT bolsters cellular resilience by ensuring that DNA lesions caused by alkylating agents are addressed promptly and effectively.

MGMT has a profound association with several types of cancer such as glioblastoma and colorectal cancer. Reduced MGMT expression or activity can lead to enhanced susceptibility to the carcinogenic effects of alkylating agents making cancer development more likely. Additionally MGMT inhibitors such as lomeguatrib are used therapeutically to enhance the effectiveness of alkylating chemotherapy. Within this context MGMT interacts with proteins involved in tumor suppression and cell cycle regulation including p53 influencing both cancer progression and treatment outcomes.

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产品实验方案

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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靶点信息

Involved in the cellular defense against the biological effects of O6-methylguanine (O6-MeG) and O4-methylthymine (O4-MeT) in DNA. Repairs the methylated nucleobase in DNA by stoichiometrically transferring the methyl group to a cysteine residue in the enzyme. This is a suicide reaction : the enzyme is irreversibly inactivated.

See full target information MGMT

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