antibodies against idh1 r132h  (Biozol Diagnostica Vertrieb GmbH)

Journal: Acta Neuropathologica Communications

Article Title: Dual phenotypes in recurrent astrocytoma, IDH-mutant; coexistence of IDH-mutant and IDH-wildtype components: a case report with genetic and epigenetic analysis

doi: 10.1186/s40478-024-01879-9

Figure Lengend Snippet: Hematoxylin and eosin (HE) staining and immunohistochemistry (IHC) of the primary and the secondary recurrent tumor. ( A ) HE staining and IHC staining using anti-IDH1 R132H antibody (inset) of the primary brain tumor. ( B ) HE staining of the second recurrent tumor. The upper area of the image shows a small round-cell tumor component (arrow). Atypical glial cells with nuclear hyperchromasia and pleomorphism are observed in the lower area (arrowhead). ( C ) IHC staining of the second recurrent tumor with anti-IDH1 R132H antibody. ( D ) IHC staining of the second recurrent tumor using anti-Ki-67 antibody. All scale bars represent 100 μm

Article Snippet: Multiple stainings were performed on the adjacent section with antibodies against IDH1 R132H (Dianova GmbH, H09 clone), GFAP (Dako, 6F2 clone), p53 (Santa Cruz Biotechnology, DO-1 clone), ATRX (Sigma-Aldrich, polyclonal), and Ki-67 (Dako, Mib-1 clone).

Techniques: Staining, Immunohistochemistry

Journal: Acta Neuropathologica Communications

Article Title: Dual phenotypes in recurrent astrocytoma, IDH-mutant; coexistence of IDH-mutant and IDH-wildtype components: a case report with genetic and epigenetic analysis

doi: 10.1186/s40478-024-01879-9

Figure Lengend Snippet: Molecular profiling of NGY-P, NGY-R1, NGY-R2A, and NGY-R2B. ( A ) A phylogeny tree depicts the optimal evolution pattern of primary and recurrent samples highlighting genes that are frequently mutated in tumors. Branch lengths are proportional to the number of mutations detected. ( B ) This panels shows the β allele frequency of chromosome 2 for each sample, where the IDH1 are located. ( C ) Kaplan–Meier curves are used to represent cases with CDKN2A homozygous deletion (HD) and PDGFR amplification (amp; n = 5), as well as cases with CDKN2A HD ( n = 9) among IDH-mutant astrocytoma. The X-axis indicates time (months) and the Y-axis indicates probability of survival. ( D ) MGMT promoter methylation status of NGY-R2A and NGY-R2B. The y-axis represents MGMT promoter methylation score, and the red dot line represents cut off value (0.3582). ( E ) t-distributed stochastic neighbor embedding (t-SNE) plot is drawn from the methylation data of our cases and reference data, which includes low-grade and high-grade gliomas. A IDH, IDH-mutant astrocytoma ( n = 78); A IDH HG, high-grade IDH-mutant astrocytoma ( n = 46); and O IDH, IDH-mutant oligodendroglioma ( n = 80). Reference methylation data for gliomas (GSE90496) were obtained from the Gene Expression Omnibus database ( http://www.ncbi.nlm.nih.gov/geo/ ). A molecular classification algorithm and copy number analysis from the German Cancer Center (DKFZ classifier, https://www.molecularneuropathology.org/mnp ) was performed . GBM, MES ( n = 56), MID ( n = 14), MYCN ( n = 16), RTK1 ( n = 64), RTK2 ( n = 143), RTK3 ( n = 13), and G34 ( n = 34) represent GBM subgroups

Article Snippet: Multiple stainings were performed on the adjacent section with antibodies against IDH1 R132H (Dianova GmbH, H09 clone), GFAP (Dako, 6F2 clone), p53 (Santa Cruz Biotechnology, DO-1 clone), ATRX (Sigma-Aldrich, polyclonal), and Ki-67 (Dako, Mib-1 clone).

Techniques: Amplification, Mutagenesis, Methylation, Expressing

Journal: Cell Death Discovery

Article Title: The development of a hiPSC-based platform to identify tissue-dependencies of IDH1 R132H

doi: 10.1038/s41420-023-01747-w

Figure Lengend Snippet: Representative Western blot membranes showing protein bands of IDH1 ( A ) and IDH1R132H ( B ), IDH1 and IDH1R132H 46 kDa, GAPDH 36 kDa, beta Actin 42 kDa. IDH1R132H protein only visible after DOX induction (12 h). Quantification of D2 Hydroxyglutarate in metabolic extracts of the corresponding cells as assessed by mass spectrometry (MS) ( C ).

Article Snippet: Blots were probed with antibody against IDH1-R132H (Dianova, DIA-H09, mouse), IDH1 (Cell Signaling Technologies, #3997, rabbit) and GFAP (ProteinTech Group Inc., #60004–1, mouse) or beta-Actin (Cell Signaling Technology, #4970, rabbit).

Techniques: Western Blot, Mass Spectrometry

Journal: Cell Death Discovery

Article Title: The development of a hiPSC-based platform to identify tissue-dependencies of IDH1 R132H

doi: 10.1038/s41420-023-01747-w

Figure Lengend Snippet: A iPS11-wt; B pSLIK-IDH1-iPS11; C pSLIK-R132H-iPS11; D pSLIK-EV-iPS11; E pSin-p53-iPS11; F pSin-p53-pSLIK-IDH1-iPS11; G : pSin-p53-pSLIK-R132H-iPS11; H : pSin-p53-pSLIK-EV-iPS11.

Article Snippet: Blots were probed with antibody against IDH1-R132H (Dianova, DIA-H09, mouse), IDH1 (Cell Signaling Technologies, #3997, rabbit) and GFAP (ProteinTech Group Inc., #60004–1, mouse) or beta-Actin (Cell Signaling Technology, #4970, rabbit).

Techniques:

Journal: Cell Death Discovery

Article Title: The development of a hiPSC-based platform to identify tissue-dependencies of IDH1 R132H

doi: 10.1038/s41420-023-01747-w

Figure Lengend Snippet: Summary of FACS-based assessment of percentage of cell expressing pluripotency markers with IDH1-cell models tested upon induction of the transgene for 48 h.

Article Snippet: Blots were probed with antibody against IDH1-R132H (Dianova, DIA-H09, mouse), IDH1 (Cell Signaling Technologies, #3997, rabbit) and GFAP (ProteinTech Group Inc., #60004–1, mouse) or beta-Actin (Cell Signaling Technology, #4970, rabbit).

Techniques: Expressing

Journal: Cell Death Discovery

Article Title: The development of a hiPSC-based platform to identify tissue-dependencies of IDH1 R132H

doi: 10.1038/s41420-023-01747-w

Figure Lengend Snippet: Global gene expression profiling revealed a distinct separation of transcriptome from cells with induced IDH1 and cells with induced IDH1R132H indicating the significance of this biomarker in the context of total gene expression networks in the context of human pluripotent stem cells ( A ). Gene Set Enrichment Analysis identified various pathways dysregulated in response to IDH1R132 induction such as increased expression of gene associated with angiogenesis or downregulation of p53 network indicating misbalanced DNA damage repair ( B ).

Article Snippet: Blots were probed with antibody against IDH1-R132H (Dianova, DIA-H09, mouse), IDH1 (Cell Signaling Technologies, #3997, rabbit) and GFAP (ProteinTech Group Inc., #60004–1, mouse) or beta-Actin (Cell Signaling Technology, #4970, rabbit).

Techniques: Expressing, Biomarker Assay

Journal: Cell Death Discovery

Article Title: The development of a hiPSC-based platform to identify tissue-dependencies of IDH1 R132H

doi: 10.1038/s41420-023-01747-w

Figure Lengend Snippet: Cell growth–drug response curves of iPS11_IDH1R132H on top three effective drugs out of a semi-automatic executed drug screening, as defined by dose-dependent reduction of cell growth reaching lowest 50% of growth inhibition concentration (IC50) when using minimal amount of drug (Plerixafor GI50 = 18.3 nM, Trametinib GI50 = 30,7 nM, Abemaciclib GI50 = 33,0 nM ( A ). Protein verification of IDH1 status of p53 mutant glial tumor models used in this study and results of drug testing on those glioblastoma cells. Trametinib and Abemaciclib show increased efficacy in cells expressing IDH1R132H protein as compared to their IDH1 WT counterparts ( B ). Microscopic images of human neural stem cell (NSC) and results of drug sensitivity testing of NSCs under induced transgene expression, showing a trend of increased resistance of R132H cells as to Trametinib and Abemaciclib as their IDH1 WT counterparts ( C ). **** p ≤ 0.0001 empty empty vector control, WT wildtype.

Article Snippet: Blots were probed with antibody against IDH1-R132H (Dianova, DIA-H09, mouse), IDH1 (Cell Signaling Technologies, #3997, rabbit) and GFAP (ProteinTech Group Inc., #60004–1, mouse) or beta-Actin (Cell Signaling Technology, #4970, rabbit).

Techniques: Inhibition, Concentration Assay, Mutagenesis, Expressing, Plasmid Preparation

Journal: Tomography

Article Title: Mutant Isocitrate Dehydrogenase 1 Expression Enhances Response of Gliomas to the Histone Deacetylase Inhibitor Belinostat

doi: 10.3390/tomography9030077

Figure Lengend Snippet: Immunoblots of lysates from the NHA and LN229 cell-line pairs (vector control (C) and mtIDH-expressing (M)) are shown. Upper blots were probed for the IDH1 R132H mutant, while the lower blots show detection of eIF5 as a loading control.

Article Snippet: Blots were probed with antibodies against IDH1 R132H mutant (MBL Life Sciences, Woburn, MA, USA), acetylated α-tubulin (Santa Cruz Biotechnology, Dallas, TX, USA), acetylated Histone-H4 (Santa Cruz), eIF5 (Santa Cruz), and GAPDH (Millipore Sigma, Burlington, MA, USA), according to the manufacturer’s recommendations.

Techniques: Western Blot, Plasmid Preparation, Expressing, Mutagenesis

Journal: Tomography

Article Title: Mutant Isocitrate Dehydrogenase 1 Expression Enhances Response of Gliomas to the Histone Deacetylase Inhibitor Belinostat

doi: 10.3390/tomography9030077

Figure Lengend Snippet: Assessment of sMRI response in GBM patients treated on our belinostat trial: ( A ) Waterfall plot for percent change in residual tumor between the pre-RT and 4-week post-RT sMRI scans, defined as volume of CNI values greater than 2. Nine patients had assessable sMRIs at both time points, comprising eight wild-type IDH GBMs (dark bars) and one mutant IDH1 GBM (light bar). Numbers above bars indicate the value of the pre-RT residual tumor volume. ( B ) The pre-RT and 4-week post-RT CNI maps for indicated slices on the 3D renderings for volumes of CNI values greater than 2 are shown for two wild-type IDH GBMs and one mutant IDH1 GBM. Arrows on the graph indicate the individual cases that are shown in this section.

Article Snippet: Blots were probed with antibodies against IDH1 R132H mutant (MBL Life Sciences, Woburn, MA, USA), acetylated α-tubulin (Santa Cruz Biotechnology, Dallas, TX, USA), acetylated Histone-H4 (Santa Cruz), eIF5 (Santa Cruz), and GAPDH (Millipore Sigma, Burlington, MA, USA), according to the manufacturer’s recommendations.

Techniques: Mutagenesis