Anti-human IDH1^R132H antibody clone H09

Antibody clone H09 reacts specifically with the isocitrate dehydrogenase 1 (IDH1) R132H point mutation in tissue sections from formalin-fixed brain tumor specimens. Heterozygous point mutations of IDH1 codon 132 are frequent in World Health Organization (WHO) grade II and III gliomas. IDH1 R132H mutations occur in approximately 70% of astrocytomas and oligodendroglial tumors.

The high frequency and distribution of the IDH1 R132H mutation among specific brain tumor entities allow the highly sensitive and specific discrimination of various tumors by immunohistochemistry, such as anaplastic astrocytoma from primary glioblastoma or diffuse astrocytoma WHO grade II from pilocytic astrocytoma or ependymoma.  Noteworthy is the discrimination of the infiltrating edge of tumors with IDH1 mutation from reactive gliosis.

This antibody is highly useful for tumor classification and in detecting single infiltrating tumor cells.

Product details & staining protocols:   IDH1 R132H in clinical diagnosis of brain tumors

A: Strong reaction of IDH1 mutation specific antibody clone H09 in tumor center of anaplastic oligoastrocytoma.
B: Infiltration zone of anaplastic astrocytoma with specific labelling of infiltrating glioma cells by antibody clone H09.
C: Identification of single tumor cells in white matter distant from tumor center with IDH1 mutation specific antibody clone H09.
D: Cortex infiltrated by Oligodendroglioma with specific labelling of tumor cells by antibody clone H09
E: Double staining of GFAP (glial fibrillary acidic protein, red) and clone H09 (brown) of oligodendroglioma infiltration zone demonstrating specific labelling of tumor cells but not of GFAP positive reactive astrocytes.
F: Strong reaction of IDH1 mutation specific antibody clone H09 with IDH1 R132H mutated diffuse astrocytoma (left) but not with wild type tumor (right).

A: Immunostainig with clone H09 (1:50 dilution) of a tissue array with different brain tumor entities: IDH1 mutation specific antibody clone H09 shows a strong reaction with IDH1 R132H mutated oligodendroglioma WHO II (R132H) but not with wild type glioblastoma (wt) or with IDH1 R132C mutated astrocytoma WHO II (IDH1 R132C).

B: diffuse astrocytoma R132C (negative) / C: diffuse astrocytoma R132H (positive) / D: glioblastoma IDH1wt (negative) / E: oligodendroglioma R132H (positive)

(pictures courtesy of Prof. Dr. Andreas von Deimling, Department of Neuropathology, University Heidelberg / Clinical Cooperation Unit Neuropathology, German Cancer Research Center (DKFZ), Heidelberg, Germany)

Instructions for use

Immunohistochemical staining of standard formalin-fixed paraffin sections
Deparaffinize and rehydrate according to standard procedures. Heat induced epitope retrival (HIER) is required.

For immunohistochemical detection different techniques can be used: Indirect immunoenzyme labeling with a secondary antibody conjugate, biotin/(strept)avidin-based detection, soluble enzyme immune complex or polymer-based detection. To detect antibody, follow the instructions provided with the particular visualization system. The antibody is suited for immuno-histochemical staining using automated platforms.

Use the antibody at 1:20 dilution for 30min at RT.

Technical note

Diffuse astrocytoma WHO grade II may have low protein-expression. At high dilution of the antibody single tumor cells in the infiltration zone may not be stained.

Anti-human IDH1 (wildtype) antibody clone W09

Rat antibody clone W09 detects wild-type IDH1 in glioblastoma and serves as control for the IDH1 point mutation specific mouse antibody clone H09.

Moreover, Tan et al. (Mol Cell Proteomics, 2011) have identified IDH1 as a potential diagnostic and prognostic biomarker for Non-small-cell Lung Cancer (NSCLC). These findings suggest that anti-IDH1 antibody could be used as a histochemical biomarker for prognosis prediction of NSCLC.

Immunohistochemistry of human IDH1 R132H and human IDH1 wild-type in formalin-fixed paraffin-embedded glioblastoma.

No reaction of IDH1 R132H mutation specific antibody clone H09 with glioblastoma (left) but strong reaction of IDH1 wild-type specific antibody clone W09 with the same glioblastoma (right). Magnification: 200x (top) and 400x (bottom)

(pictures courtesy of Prof. Dr. Andreas von Deimling, Department of Neuropathology, University Heidelberg / Clinical Co-operation Unit Neuropathology, German Cancer Research Center (DKFZ), Heidelberg, Germany)

References

1. Capper D, Zentgraf H, Balss J, Hartmann C, von Deimling A. Monoclonal antibody specific for IDH1 H132R mutation. Acta Neuropathol. 118(5): 599-601, 2009
2. Capper D, Weissert S, Balss J, Habel A, Meyer J, Jäger D, Ackermann U, Tessmer C, Korshunov A, Zentgraf H, Hartmann C, von Deimling A. Characterization of R132H mutation-specific IDH1 antibody binding in brain tumors. Brain Pathol. 20(1): 245-254, 2010
3. Andrulis M, Capper D, Luft, T, Hartmann C, Zentgraf H, von Deimling A. Detection of isocitrate dehydrogenase 1 mutation R132H in myelodysplastic syndrome by mutation-specific antibody and direct sequencing. Leuk Res. 34(8):1091-1093, 2010.
4. Andrulis M, Capper D, Meyer J, Penzel R, Hartmann C, Zentgraf H, von Deimling, A. IDH1 R132H mutation is a rare event in MPN as determinded by a mutation specific antibody. Haematologica 95(10):1797-1798, 2010
5. Sellner L, Capper D, Meyer J, Langhans CD, Hartog CM, Pfeifer H, Serve H, Ho AD, Okun JG, Krämer A, von Deimling A. Increased levels of 2-hydroxyglutarate in AML patients with IDH1-R132H and IDH2-R140Q mutations. Eur J Haematol. 85(5):457-459, 2010
6. Capper D, Sahm F, Hartmann C, Meyermann R, von Deimling A, Schittenhelm J. Application of mutant IDH1 antibody to differentiate diffuse glioma from nonneoplastic central nervous system lesions and therapy-induced changes. Am J Surg Pathol. 34(8):1199-1204, 2010
7. Horbinski C, Kofler J, Yeaney G, Carnelo-Piragua S, Venneti S, Louis DN, Perry A, Murdoch G, Nikiforova M. Isocitrat dehydrogenase 1 mutations are adverse prognostic markers in “gangliomas”. Brain Pathology 20:47, 2010
8. Hartmann C, Hentschel B, Wick W, Capper D, Felsberg J, Simon M, Westphal M, Schackert G, Meyermann R, Pietsch T, Reifenberger G, Weller W, Loeffler M, von Deimling A. Patients with IDH1 wild type anaplastic astrocytomas exhibit worse prognosis than IDH1-mutated glioblastomas, and IDH1 mutation status accounts for the unfavorable prognostic effect of higher age: implications for classification of gliomas. Acta Neuropathol. 120(6):707-718, 2010
9. Capper D, Reuss D, Schittenhelm J, Hartmann C, Bremer J, Sahm F, Harter PN, Jeibmann A, von Deimling A. Mutation-specific IDH1 antibody differntiates oligodendrogliomas and oligoastrocytomas from other brain tumors with oligodendroglioma-like morphology. Acta Neuropathol. 121(2):241-252, 2011
10. Amary MF, Bacsi K, Maggiani F, Damato S, Halai D, Berisha F, Pollock R, O'Donnell P, Grigoriadis A, Diss T, Eskandarpour M, Presneau N, Hogendoorn PC, Futreal A, Tirabosco R, Flanagan AM. IDH1 and IDH2 mutations are frequent events in central chondrosarcoma and central and periosteal chondromas but not in other mesenchymal tumours. J Pathol. 224(3):334-343, 2011
11. Sahm F, Capper D, Meyer J, Hartmann C, Herpel E, Andrulis M, Mechtersheimer G, Petersen I, Paulus W, von Deimling A. Immunohistochemical analysis of 1844 human epithelial and haematopoetic tumors and sarcomas for IDH1 R132H mutation. Histopathology 58(7):1167-1172, 2011
12. Preusser M, Wöhrer A, Stary S, Höftberger R, Streubel B, Hainfellner JA. Value and limitations of immunohistochemistry and gene sequencing for detection of the IDH1-R132H mutation in diffuse glioma biopsy specimens. J Neuropathol Exp Neurol. 70(8):715-723, 2011
13. Preusser M, Capper D, Hartmann C. IDH testing in diagnostic neuropathology: review and practical guideline article by the Euro-CNS research committee. Clinical Neuropathology 30(5):217-230, 2011
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