Name: Anti-MGMT antibody [MT3.1]
SKU: ab39253
Rating: 4 (6 reviews)

Anti-MGMT antibody [MT3.1] (ab39253) is a mouse monoclonal antibody detecting MGMT in Western Blot, IHC-P. Suitable for Human.

- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 10 publications

- Trusted since 2006

Images

Western blot - Anti-MGMT antibody [MT3.1] (AB39253), expandable thumbnail

Publications

Key facts

Isotype: IgG1
Host species: Mouse
Storage buffer: pH: 7.3 - 7.5
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.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	WB
Human	Tested	Tested
Mouse	Not recommended	Not recommended
Rat	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Associated Products

Select an associated product type

4 products for Alternative Product

Target data

See full target information MGMT

Function

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.

Alternative names

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

Key facts

Isotype: IgG1
Host species: Mouse
Storage buffer: pH: 7.3 - 7.5
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.
Clone number: MT3.1
Purification technique: Affinity purification Protein A
Concentration

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

What is this antibody validated in?

Anti-MGMT antibody [MT3.1] (ab39253) is a mouse recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunohistochemistry (IHC-P) in Human samples.

What is the molecular weight of MGMT?

Anti-MGMT [MT3.1] (ab39253) specifically detects a band for MGMT (UniProt: P16455) at a molecular weight of 22kDa.

Trusted by the scientific community

Anti-MGMT [MT3.1] (ab39253) was first used in a scientific publication in 2006 and has been cited over 10 times in peer-reviewed journals.

Reviewed by scientists

Anti-MGMT [MT3.1] (ab39253) has over 5 independent reviews from customers.

Specificity confirmed

The specificity of Anti-MGMT antibody [MT3.1] (ab39253) has been confirmed by Immunocytochemistry testing in MGMT Knockout HAP1 cells.

Other related products

We have a range of other formats of antibody clone [MT3.1] also available for your convenience:
ab39253, Carrier free - ab255966

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.

Supplementary info

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

Activity summary

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.

Associated diseases and disorders

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.

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.

7 product images

Western blot - Anti-MGMT antibody [MT3.1] (ab39253)
All lanes: Western blot - Anti-MGMT antibody [MT3.1] (ab39253) at 1/500 dilution
Lane 1: Wild type HAP1 whole cell lysate at 20 µg
Lane 2: MGMT knockout HAP1 whole cell lysate at 20 µg
Lane 3: MCF7 (human breast adenocarcinoma epithelial cell) at 20 µg
Lane 4: Jurkat (human T cell leukemia T lymphocyte at 20 µg
Predicted band size: 22 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [MT3.1] (ab39253)
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue labeling MGMT with ab39253 at 1/200 followed by a ready to use secondary. Positive staining on human prostate carcinoma. The section was incubated with ab39253 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use secondary. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
This image is courtesy of an anonymous customer review.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [MT3.1] (ab39253)
ab39253 staining MGMT in human stomach tissue sections by Immunohistochemistry. Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using 10mM citrate buffer pH6.0. Samples were then blocked with 5% serum for 1 hour at 23°C followed by incubation with the primary antibody, at a 1/100 dilution, for 1 hour at 23°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as secondary antibody.
This image was generated from the hybridoma version.
Western blot - Anti-MGMT antibody [MT3.1] (ab39253)
All lanes: Western blot - Anti-MGMT antibody [MT3.1] (ab39253) at 1/500 dilution
Lane 1: MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2: HeLa (human cervix adenocarcinoma epithelial cell ), whole cell lysate at 20 µg
Lane 3: Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg
Lane 4: HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
All lanes: Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa
Exposure time: 92s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [MT3.1] (ab39253)
Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling MGMT with ab39253 at 1/200 followed by a ready to use secondary. Positive staining on human lung carcinoma. The section was incubated with ab39253 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use secondary. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
This image is courtesy of an anonymous customer review
Western blot - Anti-MGMT antibody [MT3.1] (ab39253)
Blocking Step: 5% BSA for 1 hour at 23°C
This image was generated from the hybridoma version.
All lanes: Western blot - Anti-MGMT antibody [MT3.1] (ab39253) at 1/500 dilution
All lanes: Human OE33 cells - whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Rabbit Anti-Mouse IgG H&L (HRP) (Rabbit Anti-Mouse IgG H&L (HRP) ab6728) at 1/1000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 21 kDa
Exposure time: 30s
Western blot - Anti-MGMT antibody [MT3.1] (ab39253)
MGMT Western blot staining using mouse Anti-MGMT antibody
Western blot: Anti-MGMT antibody [MT3.1] (ab39253) staining at 1/500 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab39253 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 knockout cell line Human MGMT knockout HCT116 cell line ab286541. The band observed in the knockout 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 knockout 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-MGMT antibody [MT3.1] (ab39253) at 1/500 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: Western blot - Human MGMT knockout HCT116 cell line (Human MGMT knockout HCT116 cell line ab286541)
Lane 2: MGMT knockout HCT 116 cell lysate at 20 µg
Secondary
Lanes 1 - 2: Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 2: Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 22 kDa

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Anti-MGMT antibody [MT3.1]

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