| Field | Specification |
|---|---|
| Target | |
| Alternative names | Methylated-DNA--protein-cysteine methyltransferase;2.1.1.63;6-O-methylguanine-DNA methyltransferase;MGMT;O-6-methylguanine-DNA-alkyltransferase;MGMT; |
| UniProt # | |
| Host | |
| Clonality | |
| Isotype | |
| Reactivity | |
| Applications | |
| Immunogen | E.coli-derived human MGMT recombinant protein (Position: D2-N207). Human MGMT shares 70% and 69% amino acid (aa) sequence identity with mouse and rat MGMT, respectively. |
| Molecular weight | |
| Purification | |
| Reconstitution | |
| Cellular localization | |
| Concentration | |
| Form | Lyophilized |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Product Overview
This rabbit polyclonal antibody detects methylated-DNA--protein-cysteine methyltransferase (gene MGMT) in human samples and is validated for flow cytometry, ICC/IF, IHC and Western blot. The predicted molecular weight is 21.6 kDa, and the supplier reports an observed band at about 22–24 kDa.
O6-methylguanine-DNA methyltransferase (MGMT) is a DNA repair protein that removes methyl groups from O6-methylguanine and related lesions. It moves the methyl group onto one of its own cysteine residues, which permanently inactivates the enzyme, and it is widely studied because it counteracts DNA damage caused by alkylating agents.
| Target | Methylated-DNA--protein-cysteine methyltransferase (gene MGMT; UniProt P16455, human) |
|---|---|
| Host / Clonality / Isotype | Rabbit / Polyclonal / Rabbit IgG |
| Reactivity | Human |
| Form | Lyophilized |
| Formulation | Per vial: 0.9 mg NaCl; 0.2 mg Na2HPO4; 0.01 mg NaN3; plus stabilizers |
| Calculated MW | 21.6 kDa |
| Observed MW | 22–24 kDa |
| Storage | As supplied: −20 °C for up to 12 months from receipt. After reconstitution: 4 °C for up to 1 month, or aliquot and keep at −20 °C for up to 6 months. Avoid repeated freeze–thaw cycles. |
Validated Applications
| Western blot | 0.1–0.5 µg/mL (Human) |
|---|---|
| IHC (paraffin sections) | 2–5 µg/mL (Human) |
| Immunocytochemistry / Immunofluorescence | 0.5–1 µg/mL (Human) |
| Immunocytochemistry / Immunofluorescence | 2 µg/mL (Human) |
| Flow cytometry (fixed cells) | 1–3 µg/1×106 cells (Human) |
Samples with a confirmed band (WB): human Caco-2 cells, human Raji cells.
Recommended loading (WB): 20–40 µg of total protein per lane.
Conditions in the example images (WB): 5–20% gradient SDS-PAGE under reducing conditions, 30 µg lysate per lane, transfer to nitrocellulose membrane, blocking in 5% non-fat milk, primary antibody at 0.5 µg/mL overnight at 4 °C, HRP-conjugated secondary antibody at 1:5000, ECL detection.
Samples with confirmed staining (IHC): human colorectal cancer tissue.
Recommended antigen retrieval: heat-mediated, in TE buffer (pH 9.0); citrate buffer (pH 6.0) can be used instead.
Conditions in the example images (IHC): heat-mediated antigen retrieval in EDTA buffer (pH 8.0).
Samples with confirmed staining (ICC/IF): MCF-7 cells.
Samples with confirmed staining (ICC/IF): human MCF-7 cells.
Samples with a confirmed signal (flow cytometry): THP-1 cells.
Immunogen
Recombinant human MGMT fragment (Asp2–Asn207), expressed in E. coli. Sequence identity with the mouse and rat orthologs: 70% and 69%, respectively.
Reactivity Notes
The supplier lists reactivity with human. UniProt places methylated-DNA--protein-cysteine methyltransferase in the nucleus. The samples tested by the supplier (listed above) are a practical starting point for positive controls.
Safety
Customization & Add-ons: Can’t find the antibody you need—or require a custom format for your assay? We can help you source the best match or support custom antibody solutions for diverse research needs, including species and isotype selection, conjugations and labeling (e.g., HRP/AP, biotin, fluorophores), purification grade options (Protein A/G, affinity purified), formulation preferences (buffer selection, carrier-free, glycerol-free), custom concentrations and aliquoting, low-endotoxin options for cell-based work, and application-focused QC/validation support (project dependent). Click Talk to a Scientist to submit a request, email us at support@biohippo.com, or explore our Research Services for additional support—our team will follow up with feasibility details and next steps.
Steve Swenson et al. (2018) A Novel Venom-Derived Peptide for Brachytherapy of Glioblastoma: Preclinical Studies in Mice. Molecules. 10.3390/molecules23112918
Radu O. Minea et al. (2020) Developing a clinically relevant radiosensitizer for temozolomide-resistant gliomas. PLoS One. 10.1371/journal.pone.0238238
Qi Xiao-li et al. (2008) Expressions of MGMT and Survivin in colorectal carcinoma. Chinese Journal of Cancer Research. 10.1007/s11670-008-0139-4
