MGMT antibody (pAb)

RRID: AB_2793405
Catalog No: 39951 Format: 100 µl ¥86,000 Buy
Catalog No: 39952 Format: 10 µl ¥24,000 Buy

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Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Affinity Purified
Host:
Rabbit
Molecular Weight:
28 kDa
Reactivity:
Human

Applications

Western Blot Validated Immunoprecipitation Validated

Application Notes

Applications Validated by Active Motif:
IP: 10 µl per IP
WB*: 1:500 - 1:1,000 dilution

The addition of 0.05% Tween 20 in the blocking buffer and primary antibody incubation buffer is recommended to aid in detection by Western blot. Individual optimization may be required.

*Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western blot.

Immunogen

This MGMT antibody was raised against a peptide derived from the C-terminal region of human MGMT.

Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

 

MGMT antibody (pAb) tested by Immunoprecipitation.
10 µl of MGMT antibody was used to immunoprecipitate MGMT from 250 µg of MCF-7 nuclear cell extract (lane 2). 10 µl of rabbit IgG was used as a negative control (lane 1). The immunoprecipitated protein was detected by Western blotting using the MGMT antibody at a dilution of 1:500.

MGMT antibody (pAb) tested by Western blot.
Nuclear extract of MCF-7 cells (20 µg) probed with MGMT antibody (pAb) at a dilution of 1:500.

Background

MGMT (O-6 methylguanine DNA methyltransferase) is an enzyme involved in DNA damage repair. MGMT transfers methyl groups from O-6 methylguanine methylated DNA, and other methylated adducts, to a cysteine residue in itself, thus reversing DNA damage caused by alkylation. This is a suicide reaction as the enzyme is irreversibly inactivated. MGMT helps prevent DNA alkylation events from causing mutations. It also contributes to resistance to chemotherapy in cancer patients being treated with alkylating drugs.

Positive Control

Active Motif's MCF-7 nuclear extract (Catalog No. 36017) can be used as a positive control for MGMT antibody.

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting