Immunofluorescence analysis of MGMT (MT23.2) was performed using 70 % confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with MGMT (MT23.2) Mouse Monoclonal antibody (357000) at 2 µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominantly nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Rat, Bacteria, Human, Not Applicable|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Recombinant human MGMT|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunohistochemistry (Paraffin) (IHC (P))||See 5 publications below|
|Western Blot (WB)||See 2 publications below|
|Miscellaneous PubMed (MISC)||See 1 publications below|
|Flow Cytometry (Flow)||See 1 publications below|
|Immunohistochemistry (Paraffin, Frozen) (IHC (P, F))||See 3 publications below|
O6-Methylguanine-DNA Methyltransferase (MGMT) is an important DNA repair protein involved in tumor cell resistance to the cytostatic activity of chemotherapeutic alkylating agents. This protein is also effective in protecting normal cells against the genotoxic and carcinogenic effects of DNA alkylation. The alkylating drug resistance is caused by MGMT's ability to remove DNA alkyl groups introduced in the O6 position of guanine. MGMT is expressed in highly variable amounts, depending upon the cell and tissue type, species, and cellular growth characteristics. In addition, MGMT activity varies among groups of tumors and within a particular type of tumor.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Phase 2 study of concurrent radiotherapy and temozolomide followed by temozolomide and lomustine in the treatment of children with high-grade glioma: a report of the Children's Oncology Group ACNS0423 study.
35-7000 was used in immunohistochemistry - paraffin section to analyze the Children's Oncology Group ACNS0423 phase 2 study of concurrent temozolomide and radiotherapy followed by lomustine and temozolomide in treatment of children with high-grade glioma
|Jakacki RI,Cohen KJ,Buxton A,Krailo MD,Burger PC,Rosenblum MK,Brat DJ,Hamilton RL,Eckel SP,Zhou T,Lavey RS,Pollack IF||Neuro-oncology (18:1442)||2016|
|Not Applicable||Not Cited||
Glioma Cells in the Tumor Periphery Have a Stem Cell Phenotype.
35-7000 was used in immunohistochemistry - paraffin section to study tumor periphery glioma cells and their stem cell phenotype
|Munthe S,Petterson SA,Dahlrot RH,Poulsen FR,Hansen S,Kristensen BW||PloS one (11:null)||2016|
Long-term outcome and MGMT as a predictive marker in 24 patients with atypical pituitary adenomas and pituitary carcinomas given treatment with temozolomide.
35-7000 was used in immunohistochemistry - paraffin section to discuss the use of temozolomide to treat pituitary tumors
|Bengtsson D,Schrøder HD,Andersen M,Maiter D,Berinder K,Feldt Rasmussen U,Rasmussen ÅK,Johannsson G,Hoybye C,van der Lely AJ,Petersson M,Ragnarsson O,Burman P||The Journal of clinical endocrinology and metabolism (100:1689)||2015|
Impact of morphology, MIB-1, p53 and MGMT on outcome in pilocytic astrocytomas.
35-7000 was used in immunohistochemistry - paraffin section to discuss indicators of tumor aggressiveness for pilocytic astrocytoma
|Horbinski C,Hamilton RL,Lovell C,Burnham J,Pollack IF||Brain pathology (Zurich, Switzerland) (20:581)||2010|
|Not Applicable||Not Cited||
Prognostic significance of DNA repair proteins MLH1, MSH2 and MGMT expression in non-small-cell lung cancer and precursor lesions.
35-7000 was used in immunohistochemistry - paraffin section to investigate the role of DNA repair proteins and their prognostic significance in non-small-cell lung cancer
|Cooper WA,Kohonen-Corish MR,Chan C,Kwun SY,McCaughan B,Kennedy C,Sutherland RL,Lee CS||Histopathology (52:613)||2008|
Implication of a Chromosome 15q15.2 Locus in Regulating UBR1 and Predisposing Smokers to MGMT Methylation in Lung.
35-7000 was used in western blot to show that UBR regulates MGMT homeostasis and DNA repair of alkylated DNA adducts in cells.
|Leng S,Wu G,Collins LB,Thomas CL,Tellez CS,Jauregui AR,Picchi MA,Zhang X,Juri DE,Desai D,Amin SG,Crowell RE,Stidley CA,Liu Y,Swenberg JA,Lin Y,Wathelet MG,Gilliland FD,Belinsky SA||Cancer research (75:3108)||2015|
Enhanced MGMT expression contributes to temozolomide resistance in glioma stem-like cells.
35-7000 was used in western blot to study the role of MGMT in the resistance against temozolomide in glioma stem-like cells.
|Qiu ZK,Shen D,Chen YS,Yang QY,Guo CC,Feng BH,Chen ZP||Chinese journal of cancer (33:115)||2014|
Hsp27 (HSPB1): a possible surrogate molecular marker for loss of heterozygosity (LOH) of chromosome 1p in oligodendrogliomas but not in astrocytomas.
35-7000 was used in immunohistochemistry (paraffin) to examine heat shock proteins in different subtypes of gliomas.
|Castro GN,Cayado-Gutiérrez N,Moncalero VL,Lima P,De Angelis RL,Chávez V,Cuello-Carrión FD,Ciocca DR||Cell stress and chaperones (17:779)||2012|
|Not Applicable||Not Cited||
O6-Methylguanine-DNA methyltransferase protein expression by immunohistochemistry in brain and non-brain systemic tumours: systematic review and meta-analysis of correlation with methylation-specific polymerase chain reaction.
35-7000 was used in flow cytometry and immunohistochemistry to review methods to analyze O6-Methylguanine-DNA methyltransferase
|Brell M,Ibáñez J,Tortosa A||BMC cancer (11:null)||2011|
|Silencing of MGMT expression by promoter hypermethylation in the metaplasia-dysplasia-carcinoma sequence of Barrett's esophagus.||Kuester D,El-Rifai W,Peng D,Ruemmele P,Kroeckel I,Peters B,Moskaluk CA,Stolte M,Mönkemüller K,Meyer F,Schulz HU,Hartmann A,Roessner A,Schneider-Stock R||Cancer letters (275:117)||2009|
|Human||Not Cited||O6-methylguanine-DNA methyltransferase expression strongly correlates with outcome in childhood malignant gliomas: results from the CCG-945 Cohort.||Pollack IF,Hamilton RL,Sobol RW,Burnham J,Yates AJ,Holmes EJ,Zhou T,Finlay JL||Journal of clinical oncology : official journal of the American Society of Clinical Oncology (24:3431)||2006|
|Promoter hypermethylation of p16INK4a, E-cadherin, O6-MGMT, DAPK and FHIT in adenocarcinomas of the esophagus, esophagogastric junction and proximal stomach.||Schildhaus HU,Kröckel I,Lippert H,Malfertheiner P,Roessner A,Schneider-Stock R||International journal of oncology (26:1493)||2005|