MGMT Antibody (MT 23.2) - Azide and BSA Free

Western Blot: MGMT Antibody (MT 23.2) - Azide and BSA Free [NBP2-80847] - Detection of MGMT in MCF7 lysate using NB100-168 at 1:1000 dilution. Image from the standard format of this antibody.

Immunocytochemistry/Immunofluorescence: MGMT Antibody (MT 23.2) - Azide and BSA Free [NBP2-80847] - MCF7 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-MGMT (MT 23.2) NB100-168 at a 1:200 dilution overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was counterstained with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective. Image from the standard format of this antibody.

Immunohistochemistry-Paraffin: MGMT Antibody (MT 23.2) - Azide and BSA Free [NBP2-80847] - IHC analysis of a formalin fixed paraffin-embedded (FFPE) human kidney using 1:2000 conc. of MGMT antibody (clone MT 23.2) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodiu

Flow Cytometry: MGMT Antibody (MT 23.2) - Azide and BSA Free [NBP2-80847] - Detection of MGMT in Human Jurkat Cell Line by Flow Cytometry. Human Jurkat cell line was stained with Mouse Anti- MGMT Monoclonal Antibody (Catalog # NB100-168, filled histogram), or Mouse IgG2B isotype control (Catalog # MAB0041, open histogram) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a Bio-Techne brand. Image from the standard format of this antibody.

Immunocytochemistry/Immunofluorescence: MGMT Antibody (MT 23.2) - Azide and BSA Free [NBP2-80847] - Detection of MGMT (Green) in Hela cells using NB100-168 at a 1:50 dilution. Nuclei (Blue) are counterstained using Hoechst 33258. Image from the standard format of this antibody.

Flow Cytometry: MGMT Antibody (MT 23.2) - Azide and BSA Free [NBP2-80847] - An intracellular stain was performed on HeLa cells with MGMT Antibody (MT 23.2) antibody NB100-168 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) PE-conjugated secondary antibody (F0102B, R&D Systems). Image from the standard format of this antibody.

Simple Western: MGMT Antibody (MT 23.2) - Azide and BSA Free [NBP2-80847] - Simple Western lane view shows a specific band for MGMT in 0.2 mg/ml of MCF-7 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Image from the standard format of this antibody.

• Reactivity: Hu, Mu(-)
• Applications: WB, Simple Western, Flow, ICC/IF, IHC, CyTOF-ready
• Clone: MT 23.2
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: 1 mg/ml

MGMT Antibody (MT 23.2) - Azide and BSA Free Summary

• Description: Novus Biologicals Mouse MGMT Antibody (MT 23.2) - Azide and BSA Free (NB100-168) is a monoclonal antibody validated for use in IHC, WB, Flow, ICC/IF and Simple Western. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: Recombinant human MGMT (O6-methylguanine-DNA methyltransferase) purified from E. coli [UniProt# P16455]
• Localization: Nuclear
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: MGMT
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow (Intracellular)
  • Flow Cytometry 1-2 ug/ml
  • Immunocytochemistry/ Immunofluorescence 1:500
  • Immunohistochemistry 1:500-1000
  • Immunohistochemistry-Paraffin 1:500-1:1000
  • Simple Western 1:1000
  • Western Blot 1:1000
• Application Notes: This MGMT (MT 23.2) antibody is useful for Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry on paraffin-embedded sections and Western Blot. In WB, a band at approx. 24 kDa can be seen. In ICC/IF, strong staining has been seen in CEM cells and no signal has been seen in TK6 cells.
  
  In Simple Western only 10 - 15 ul of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: antibody dilution of 1:1000. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Theoretical MW: 24 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

This antibody is mouse reactivity negative.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: No Preservative
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for MGMT Antibody (MT 23.2) - Azide and BSA Free

• 6-O-methylguanine-DNA methyltransferase
• EC 2.1.1.63
• methylated-DNA--protein-cysteine methyltransferase
• methylguanine-DNA methyltransferase
• MGMT
• O-6-methylguanine-DNA methyltransferase
• O6-methylguanine-DNA methyltransferase
• O-6-methylguanine-DNA-alkyltransferase

Background

MGMT (6-O-methylguanine-DNA methyltransferase) is a DNA repair enzyme with the ability to protect cells from carcinogenic effects of alkylating agents by removing adducts from the O(6) position of guanine. MGMT repairs alkylated guanine residues in DNA by stoichiometrically transferring the alkyl group at the O-6 position to a cysteine residue in the enzyme, and in this so called 'suicidal reaction', the enzyme is irreversibly inactivated. Localized in the nucleus, MGMT is ubiquitously expressed in normal human tissues with overexpression observed in several types of cancerous cells wherein it provides resistance against alkylating drugs. However, it is found epigenetically silenced in some cancers that include lymphoma, breast and prostate cancer, as well as retinoblastoma, wherein this silencing is thought to be due to promoter methylation which interferes with transcriptional factors binding. MGMT silencing has been linked to mutations in other tumor-related genes including silencing of p53 (non-small cell lung cancer and astrocytic tumors), k-ras gene mutations (gastric and colorectal cancer), and methylation of CDKN1A gene (encodes p21) as well as CDKN2A gene (encodes p16).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: MGMT