MTAP Antibody (1113601) [Unconjugated]

Western Blot shows lysates of HT‑29 human colon adenocarcinoma cell line, HCT‑116 human colorectal carcinoma cell line and Raji human Burkitt's lymphoma cell line. PVDF membrane was probed with 1 µg/ml of Mouse Anti-Human MTAP Monoclonal Antibody (Catalog # MAB11747) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for MTAP at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

MTAP was detected in immersion fixed paraffin-embedded sections of human colon using Mouse Anti-Human MTAP Monoclonal Antibody (Catalog # MAB11747) at 5 µg/ml overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using the HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm and nucleus. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

MTAP was detected in immersion fixed paraffin-embedded sections of human kidney using Mouse Anti-Human MTAP Monoclonal Antibody (Catalog # MAB11747) at 5 µg/ml overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using the HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm and nucleus. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

MTAP was detected in immersion fixed HT‑29 human colon adenocarcinoma cell line using Mouse Anti-Human MTAP Monoclonal Antibody (Catalog # MAB11747) at 8 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm and nucleus. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

MTAP was detected in immersion fixed MCF‑7 human breast cancer cell line using Mouse Anti-Human MTAP Monoclonal Antibody (Catalog # MAB11747) at 8 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm and nucleus. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

• Reactivity: Hu
• Applications: WB, IHC, ICC/IF
• Clone: 1113601
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

MTAP Antibody (1113601) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human MTAP
  Accession # Q13126
• Specificity: Detects recombinant human MTAP protein in Direct ELISA
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3-25 ug/mL
  • Immunohistochemistry 3-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for MTAP Antibody (1113601) [Unconjugated]

• 5'-methylthioadenosine phosphorylase
• BDMF
• C86fus
• DMSFH
• DMSMFH
• EC 2.4.2.28
• HEL-249
• LGMBF
• methylthioadenosine phosphorylase
• MSAPMeSAdo phosphorylase
• MTA phosphorylase
• MTAP
• MTAPase
• S-methyl-5'-thioadenosine phosphorylase

Background

Methyl-thioadenosine phosphorylase/MTAP is part of the PNP/MTAP phosphorylase family and catalyzes the reversible phosphorylation of S-methyl-5'-thioadenosine (MTA), a major byproduct of polyamine synthesis essential for cell growth and proliferation. MTAP also produces most of the free adenine generated in human cells through a salvage pathway and thus couples the purine salvage pathway with polyamine biosynthesis. MTAP forms an active trimer where each identical 32 kDa monomer contains a separate active site (1). Each active site contains three distinct regions required for base-, methylthioribose-, and sulfate/phosphate-binding (1). MTAP is cytosolic and abundantly expressed in normal cells and tissues (2). In contrast, deficient MTAP expression is observed in many types of tumors including lung, bladder, pancreatic, and endometrial cancer (3) due to hyper-methylation gene suppression (4) or gene deletion (3, 5, 6). MTA accumulation leads to an immunosuppressive tumor microenvironment and apoptotic resistance (7-9) and MTAP directly regulates the level of MTA present. MTAP has been reported as a tumor suppressor (6,10) that may also act in a manner that is independent of enzymatic activity (11) through signaling pathways such as the insulin-like growth factor-1 receptor pathway (12). Potential therapeutic strategies to exploit MTAP deficiency in tumors (13,14) or inhibit MTAP in tumors that express MTAP, such as prostate cancer, are under investigation (15).

1. Appleby, T.C. et al. (1999) Structure 7:629.
2. Garbers, D.L. (1978) Biochim. Biophys. Acta. 523:82.
3. Li, Y. et al. (2019) J. Cancer 10:927.
4. Hellerbrand, C. et al. (2006) Carcinogenesis 27:64.
5. Cairns, P. et al. (1995) Nat. Genet. 11:210.
6. Christopher, S.A. (2002) Cancer Res. 62:6639.
7. Kirovski, G. et al. (2011) Am. J. Pathol. 178:1145.
8. Czech, B. et al. (2013) PLoS One 8:e80703.
9. Henrich, F.C. et al. (2016) Oncoimmunology 5:e1184802.
10. Kadariya, Y. et al. (2009) Cancer Res. 69:5961.
11. Tang, B. et al. (2014) G3 (Bethesda) 5:35.
12. Xu, J. et al. (2019) Signal Transduct. Target Ther. 4:2.
13. Bertino, J.R. et al. (2011) Cancer Biol. Ther. 11:627.
14. Tang, B. et al. (2018) Cancer Res. 78:4386.
15. Bistulfi, G. et al. (2016) Oncotarget 7:14380.

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.