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Purified Mouse Anti-Human MGMT
Purified Mouse Anti-Human MGMT
Immunoprecipitation (IP) and Western Blot analysis (WB) of MGMT in MOLT-4 cells (Left Panel).  MOLT-4 (Human T-lymphoblasts; ATCC CRL-1582) cell lysates were immunoprecipitated with the Mouse Anti-Human MGMT antibody (lane 1) or with an isotype control (lane 2). MGMT is identified as an ~25 kD band. The bands above the 25 kD MGMT band represent the heavy and light chains of the immunoprecipitating antibody. Western blot analysis of MGMT in MOLT-4 cell lysates probed with the Mouse Anti-Human MGMT antibody (lane 3) or an isotype control (lane 4).   Immunocytochemistry for MGMT (Middle and Right Panels). Cytospins of HeLa cells (Human cervical epitheloid carcinoma; ATCC CCL-2) were acetone-fixed and stained with the purified Mouse Anti-Human MGMT antibody (middle panel) or with an isotype control (right panel). MGMT staining is localized to the nucleus of the cells.
Immunoprecipitation (IP) and Western Blot analysis (WB) of MGMT in MOLT-4 cells (Left Panel).  MOLT-4 (Human T-lymphoblasts; ATCC CRL-1582) cell lysates were immunoprecipitated with the Mouse Anti-Human MGMT antibody (lane 1) or with an isotype control (lane 2). MGMT is identified as an ~25 kD band. The bands above the 25 kD MGMT band represent the heavy and light chains of the immunoprecipitating antibody. Western blot analysis of MGMT in MOLT-4 cell lysates probed with the Mouse Anti-Human MGMT antibody (lane 3) or an isotype control (lane 4).   Immunocytochemistry for MGMT (Middle and Right Panels). Cytospins of HeLa cells (Human cervical epitheloid carcinoma; ATCC CCL-2) were acetone-fixed and stained with the purified Mouse Anti-Human MGMT antibody (middle panel) or with an isotype control (right panel). MGMT staining is localized to the nucleus of the cells.
Product Details
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BD Pharmingen™
Methylguanine-DNA Methyltransferase
Human (QC Testing)
Mouse IgG1, κ
Western blot (Routinely Tested), Immunoprecipitation, Intracellular staining (flow cytometry) (Tested During Development)
21-25 kDa
0.5 mg/ml
AB_396564
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. An isotype control should be used at the same concentration as the antibody of interest.
557045 Rev. 5
Antibody Details
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MT5.1

The repair of mismatched DNA is essential to maintaining the integrity of genetic information over time. The important role played by DNA repair enzymes is emphasized by the fact that they are highly conserved from bacteria to yeast to mammals. In bacteria, DNA repair involves the activity of repair enzymes including the MutL, MutH, and MutS proteins, as well as an enzyme originally identified in bacteria, and subsequently in all mammalian species, MGMT (methylguanine-DNA methyltransferase). Human MGMT is a 21 kDa protein which protects cells from the mutagenic effect of alkylating agents, which add a methyl group to the O6 position of guanine in DNA, producing O6-methylguanine. While MGMT is generally ubiquitously expressed in mammalian cells, the expression levels within cell and tissue types can be quite variable. Certain tumor cell types have been identified which express neither MGMT protein or mRNA, despite the apparent normal expression of the MGMT gene. The variable expression of MGMT may be a factor in the susceptibility of cells and tissues to the mutagenic effects of alkylating agents. It has been suggested that methylation of cytosine in the MGMT gene and promoter region may correlate with the expression of the MGMT gene product.

557045 Rev. 5
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
557045 Rev.5
Citations & References
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Development References (5)

  1. Ayi TC, Loh KC, Ali RB, Li BF. Intracellular localization of human DNA repair enzyme methylguanine-DNA methyltransferase by antibodies and its importance. Cancer Res. 1992; 52(23):6423-6430. (Biology). View Reference
  2. Fornace AJ Jr, Papathanasiou MA, Hollander MC, Yarosh DB. Expression of the O6-methylguanine-DNA methyltransferase gene MGMT in MER+ and MER- human tumor cells. Cancer Res. 1990; 50(24):7908-7911. (Biology). View Reference
  3. Gerson SL, Trey JE, Miller K, Berger NA. Comparison of O6-alkylguanine-DNA alkyltransferase activity based on cellular DNA content in human, rat and mouse tissues. Carcinogenesis. 1986; 7(5):745-749. (Biology). View Reference
  4. Woodhead AD, Grist E, Carlson C, White TE, Waldstein E, Cao EH. Presence of O6-methylguanine acceptor protein in the tissues of different classes of vertebrates and invertebrates. Comp Biochem Physiol B Biochem Mol Biol. 1986; 85(1):125-130. (Biology). View Reference
  5. von Wronski MA, Brent TP. Effect of 5-azacytidine on expression of the human DNA repair enzyme O6-methylguanine-DNA methyltransferase. Carcinogenesis. 1994; 15(4):577-582. (Biology). View Reference
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557045 Rev. 5

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.