Human MGMT ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human MGMT in Cell Lysate, Tissue Extracts samples.
Images
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell Lysate, Tissue Extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 1.94 - 62 ng/mL
- Assay time
- 1h 30m
- Sensitivity
- = 0.55 ng/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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
What's included?
Recommended products
Human MGMT ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human MGMT in Cell Lysate, Tissue Extracts samples.
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell Lysate, Tissue Extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 1.94 - 62 ng/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 0.55 ng/mL
- General recovery
- 113%
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Extract | n 8 | mean - | SD - | C.V. 3.2 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Extract | n 3 | mean - | SD - | C.V. 6.6 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell Lysate | Average % = 113 | Range 107 - 122 % |
Sample type Tissue Extracts | Average % = 119 | Range 115 - 123 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
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.
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.
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.
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4 product images
Sandwich ELISA - Human MGMT ELISA Kit (ab320053)
Example of human MGMT standard curve. Background-subtracted data values (mean +/- SD) are graphed.
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.
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.
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.
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Product protocols
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