AB320053

Human MGMT ELISA Kit

Recombinant
SimpleStep
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Human MGMT ELISA Kit is a single-wash 90-min Simplestep used to quantify Human MGMT with a sensitivity of 0.55 ng/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 10x96 plates for bulk orders and 384-well plate for higher throughput
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

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

5 Images

Sandwich ELISA - Human MGMT ELISA Kit (AB320053)

sELISA

Supplier Data

Sandwich ELISA - Human MGMT ELISA Kit (AB320053)

Interpolated concentration of native MGMT was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 500 µg/mL Jurkat cell extract. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made 1X Cell Extraction Buffer + Enhancer.

sELISA

Supplier Data

Sandwich ELISA - Human MGMT ELISA Kit (AB320053)

Interpolated concentration of native MGMT was measured in duplicate at different sample concentrations. Undiluted samples are as follows : Jurkat extract 500 µg/mL, MOLT-4 extract 1,000 µg/mL, SH-SY5Y extract 1,000 µg/mL, and lung tissue extract 1,000 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer + Enhancer.

sELISA

Supplier Data

Sandwich ELISA - Human MGMT ELISA Kit (AB320053)

Example of human MGMT standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.

sELISA

Supplier Data

Sandwich ELISA - Human MGMT ELISA Kit (AB320053)

Example of human MGMT standard curve. Background-subtracted data values (mean +/- SD) are graphed.

sELISA

Supplier Data

Sandwich ELISA - Human MGMT ELISA Kit (AB320053)

Representative image of Human MGMT ELISA Kit (ab320053)

Components shown from left to right :

- 10X Wash Buffer PT (ab206977)

- 10X Human MGMT ELISA Kit Capture Antibody

- 10X Human MGMT Detector Antibody

- 10X Human MGMT ELISA Kit Lyophilised Purified Protein (2 vials)

- Human MGMT ELISA Kit 50X Cell Extraction Enhancer Solution (ab193971)

- Sample diluent NS

- TMB solution

- Stop Solution

- 5X Cell Extraction Buffer PTR (ab193970)

- Antibody Diluent 4BR

SimpleStep Pre-Coated 96-Well Microplate (ab206978) - shown behind bottles / vials.

Plate Seals - shown underneath bottles / vials.

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the ELISA kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate, Tissue Extracts

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 0.55 ng/mL

Range

1.94 - 62 ng/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

Human MGMT SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of MGMT protein in Cell Lysate and Tissue Extracts. Quantitate Human MGMT with 0.55 ng/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips, available in 10-pack (10 x 96-well plates)
-Also available in a fully validated 384-well format.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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Precision

[ { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "3.2" }, { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "6.6" } ]

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Recovery

[ { "sample": "Cell Lysate", "range": "107 - 122 %", "average": "= 113" }, { "sample": "Tissue Extracts", "range": "115 - 123 %", "average": "= 119" } ]

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What's included?

{ "values": { "1x384Tests": { "sellingSize": "1 x 384 Tests", "publicAssetCode":"ab320053-1x384Tests", "assetComponentDetails": [ { "size":"1 x 50 mL", "name":"Cell Extraction Buffer SSW", "number":"AB320053-CMP11" }, { "size":"1 x 5 mL", "name":"50X Cell Extraction Enhancer Solution (ab193971)", "number":"AB320053-CMP13" }, { "size":"2 x 12 mL", "name":"Stop Solution", "number":"AB320053-CMP08" }, { "size":"2 x 12 mL", "name":"TMB Development Solution", "number":"AB320053-CMP07" }, { "size":"1 x 384 Unit", "name":"384 well CaptSure™ microplates", "number":"AB320053-CMP12" }, { "size":"1 x 600 µL", "name":"10X Human MGMT Detector Antibody", "number":"AB320053-CMP02" }, { "size":"1 x 600 µL", "name":"10X Human MGMT Capture Antibody", "number":"AB320053-CMP01" }, { "size":"1 x 1 Unit", "name":"Plate Seal", "number":"AB320053-CMP10" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB320053-CMP05" }, { "size":"2 x 1 Vial", "name":"Human MGMT Lyophilized Recombinant Protein", "number":"AB320053-CMP03" }, { "size":"1 x 6 mL", "name":"Antibody Diluent 4BR", "number":"AB320053-CMP04" } ] }, "10x96Tests": { "sellingSize": "10 x 96 Tests", "publicAssetCode":"ab320053-10x96Tests", "assetComponentDetails": [ { "size":"1 x 50 mL", "name":"Cell Extraction Buffer SSW", "number":"AB320053-CMP11" }, { "size":"10 x 1 mL", "name":"50X Cell Extraction Enhancer Solution (ab193971)", "number":"AB320053-CMP13" }, { "size":"1 x 120 mL", "name":"Stop Solution", "number":"AB320053-CMP08" }, { "size":"1 x 120 mL", "name":"TMB Development Solution", "number":"AB320053-CMP07" }, { "size":"10 x 96 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB320053-CMP09" }, { "size":"10 x 6 mL", "name":"Antibody Diluent 4BR", "number":"AB320053-CMP04" }, { "size":"10 x 600 µL", "name":"10X Human MGMT Capture Antibody", "number":"AB320053-CMP01" }, { "size":"1 x 200 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB320053-CMP05" }, { "size":"10 x 1 Unit", "name":"Plate Seal", "number":"AB320053-CMP10" }, { "size":"20 x 1 Vial", "name":"Human MGMT Lyophilized Recombinant Protein", "number":"AB320053-CMP03" }, { "size":"10 x 600 µL", "name":"10X Human MGMT Detector Antibody", "number":"AB320053-CMP02" } ] }, "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab320053-1x96Tests", "assetComponentDetails": [ { "size":"1 x 10 mL", "name":"Cell Extraction Buffer SSW", "number":"AB320053-CMP11", "productcode":"" }, { "size":"1 x 12 mL", "name":"Stop Solution", "number":"AB320053-CMP08", "productcode":"" }, { "size":"1 x 1 mL", "name":"50X Cell Extraction Enhancer Solution (ab193971)", "number":"AB320053-CMP13", "productcode":"" }, { "size":"1 x 12 mL", "name":"Sample Diluent NS (ab193972)", "number":"AB320053-CMP06", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB Development Solution", "number":"AB320053-CMP07", "productcode":"" }, { "size":"1 x 96 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB320053-CMP09", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human MGMT Capture Antibody", "number":"AB320053-CMP01", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human MGMT Detector Antibody", "number":"AB320053-CMP02", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human MGMT Lyophilized Recombinant Protein", "number":"AB320053-CMP03", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB320053-CMP05", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seal", "number":"AB320053-CMP10", "productcode":"" }, { "size":"1 x 6 mL", "name":"Antibody Diluent 4BR", "number":"AB320053-CMP04", "productcode":"" } ] } } }

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Properties and storage information

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

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Supplementary information

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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Product protocols

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

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Target data

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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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

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