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Rabbit Recombinant Monoclonal MGMT antibody. Carrier free. Suitable for IHC-P, WB and reacts with Human samples.

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Images

Western blot - Anti-MGMT antibody [EPR4397] - BSA and Azide free (AB233012), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [EPR4397] - BSA and Azide free (AB233012), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [EPR4397] - BSA and Azide free (AB233012), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [EPR4397] - BSA and Azide free (AB233012), expandable thumbnail
  • Western blot - Anti-MGMT antibody [EPR4397] - BSA and Azide free (AB233012), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBIPICC/IFFlow Cyt
Human
Tested
Tested
Not recommended
Not recommended
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
-
Notes

Antigen retrieval is recommended.

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse
Dilution info
-
Notes

-

Associated Products

Select an associated product type

3 products for Alternative Product

Target data

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

Recommended products

Rabbit Recombinant Monoclonal MGMT antibody. Carrier free. Suitable for IHC-P, WB and reacts with Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR4397
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab233012 is the carrier-free version of Anti-MGMT antibody [EPR4397] ab108630.

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

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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.

5 product images

  • Western blot - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012), expandable thumbnail

    Western blot - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012)

    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 - unpurified Anti-MGMT antibody [EPR4397] ab108630 observed at 22 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    Anti-MGMT antibody [EPR4397] ab108630 was shown to specifically react with MGMT when MGMT knockout samples were used. Wild-type and MGMT knockout samples were subjected to SDS-PAGE. Anti-MGMT antibody [EPR4397] ab108630 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MGMT antibody [EPR4397] ab108630).

    All lanes: Western blot - Anti-MGMT antibody [EPR4397] (Anti-MGMT antibody [EPR4397] ab108630)

    Predicted band size: 22 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012)

    Anti-MGMT antibody [EPR4397] ab108630, at a 1/250 dilution, staining MGMT in paraffin embedded Human clear cell carcinoma tissue by Immunohistochemistry.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MGMT antibody [EPR4397] ab108630).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung cancer tissue sections labeling MGMT with purified Anti-MGMT antibody [EPR4397] ab108630 at 1:800 dilution (0.94 μg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MGMT antibody [EPR4397] ab108630)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012)

    Anti-MGMT antibody [EPR4397] ab108630, at a 1/250 dilution, staining MGMT in paraffin embedded Human spleen tissue by Immunohistochemistry.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MGMT antibody [EPR4397] ab108630).

  • Western blot - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012), expandable thumbnail

    Western blot - Anti-MGMT antibody [EPR4397] - BSA and Azide free (ab233012)

    This data was developed using Anti-MGMT antibody [EPR4397] ab108630, the same antibody clone in a different buffer formulation.

    Western blot: Anti-MGMT antibody [EPR4397] (Anti-MGMT antibody [EPR4397] ab108630) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-MGMT antibody [EPR4397] ab108630 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 Human MGMT knockout HCT116 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 [EPR4397] (Anti-MGMT antibody [EPR4397] ab108630) 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 (Human MGMT knockout HCT116 cell line ab286541)

    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

    Performed under reducing conditions.

    Observed band size: 22 kDa

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Product protocols

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

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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