rabbit polyclonal anti mad2 antibody  (Proteintech)

Journal: Cancer Gene Therapy

Article Title: BAG2 , MAD2L1 , and MDK are cancer-driver genes and candidate targets for novel therapies in malignant pleural mesothelioma

doi: 10.1038/s41417-024-00805-4

Figure Lengend Snippet: Scores of the signature genes resulted from the differential expression analysis using as input the EGA RNA-seq data compared to the three lung samples.

Article Snippet: IHC was carried out on 3.5 μm paraffin sections by using recombinant Anti-BAG2 Rabbit mAb [EPR3567] (1:400), Anti-Midkine Rabbit mAb [EP1143Y] (1:100; cod ab79406, ab52637; Abcam, Cambridge, MA, USA), and Anti-MAD2 (MAD2L1) Rabbit pAb (1:250; cod TA308923; OriGene, Rockville, MD, USA).

Techniques: Expressing

Journal: Cancer Gene Therapy

Article Title: BAG2 , MAD2L1 , and MDK are cancer-driver genes and candidate targets for novel therapies in malignant pleural mesothelioma

doi: 10.1038/s41417-024-00805-4

Figure Lengend Snippet: A The blot of siRNA BAG2 and MDK, C the blot of siRNA MAD2L1. β-Tubulin was used as a loading control and GAPDH was the positive control of the siRNA transfection. As shown in the blots and also in graphs ( B ) and ( D ) there was a significant reduction of the protein level in the silenced cells, as compared to the negative CTRL. E Graph showing the comparison of protein expression in MeT-5A and MSTO cell lines of BAG2, MAD2L1, and MDK (Mann–Whitney test; p value < 0.05 were considered significant, p value = 0.0332(*), 0.0021(**), 0.0002(***), <0.0001(****)).

Article Snippet: IHC was carried out on 3.5 μm paraffin sections by using recombinant Anti-BAG2 Rabbit mAb [EPR3567] (1:400), Anti-Midkine Rabbit mAb [EP1143Y] (1:100; cod ab79406, ab52637; Abcam, Cambridge, MA, USA), and Anti-MAD2 (MAD2L1) Rabbit pAb (1:250; cod TA308923; OriGene, Rockville, MD, USA).

Techniques: Control, Positive Control, Transfection, Comparison, Expressing, MANN-WHITNEY

Journal: Cancer Gene Therapy

Article Title: BAG2 , MAD2L1 , and MDK are cancer-driver genes and candidate targets for novel therapies in malignant pleural mesothelioma

doi: 10.1038/s41417-024-00805-4

Figure Lengend Snippet: A AOC of MeT-5A and MSTO at 72 h after treatment with siRNA-negative-control (siRNA CTRL−), siRNA BAG2, siRNA MAD2L1, siRNA MDK. The AOC is normalized to the time of transfection ( t 0 ). B Overall, FI of MeT-5A and MSTO was measured at 72 h after silencing; the graphs show the comparison between MeT-5A and MSTO for the three-siRNA transfection. ANOVA test; p value = 0.0332(*), 0.0021(**), 0.0002(***), <0.0001(****). C – E End point of apoptosis analysis in MSTO and MeT-5A cells in the presence of 5 µM caspase 3/7 Green Dye and siRNA BAG2, siRNA MAD2L1, and siRNA MDK, respectively. Mann–Whitney test; p value 0.0332(*), 0.0021(**), 0.0002(***), <0.0001(****).

Article Snippet: IHC was carried out on 3.5 μm paraffin sections by using recombinant Anti-BAG2 Rabbit mAb [EPR3567] (1:400), Anti-Midkine Rabbit mAb [EP1143Y] (1:100; cod ab79406, ab52637; Abcam, Cambridge, MA, USA), and Anti-MAD2 (MAD2L1) Rabbit pAb (1:250; cod TA308923; OriGene, Rockville, MD, USA).

Techniques: Negative Control, Transfection, Comparison, MANN-WHITNEY

Journal: Cancer Gene Therapy

Article Title: BAG2 , MAD2L1 , and MDK are cancer-driver genes and candidate targets for novel therapies in malignant pleural mesothelioma

doi: 10.1038/s41417-024-00805-4

Figure Lengend Snippet: A – D Representative BAG2 immunostainings in RMP, EMM, BMM, and SMM, respectively. Moderate to strong expression in the three types of MPM, while no expression is visible in the hyperplastic mesothelium in RMP (red arrow). E – H Representative MAD2L1 immunostainings in RMP, EMM, BMM, and SMM, respectively. Strong expression is present in the three types of MPM, but there are areas of discernable expression in the mesothelial cells on the pleural surface in RMP as well (blue arrow positive mesothelial area, red arrow negative area). I – L Representative MDK immunostainings in RMP, EMM, BMM, and SMM, respectively. Strong expression is seen in the three types of MPM; however, some focal expression is also detectable in the mesothelial cells on the pleural surface in RMP (blue arrow focal staining, red arrow negative staining). Magnification: RMPs ×400, MPMs ×200.

Article Snippet: IHC was carried out on 3.5 μm paraffin sections by using recombinant Anti-BAG2 Rabbit mAb [EPR3567] (1:400), Anti-Midkine Rabbit mAb [EP1143Y] (1:100; cod ab79406, ab52637; Abcam, Cambridge, MA, USA), and Anti-MAD2 (MAD2L1) Rabbit pAb (1:250; cod TA308923; OriGene, Rockville, MD, USA).

Techniques: Expressing, Staining, Negative Staining

Journal: Cancer Gene Therapy

Article Title: BAG2 , MAD2L1 , and MDK are cancer-driver genes and candidate targets for novel therapies in malignant pleural mesothelioma

doi: 10.1038/s41417-024-00805-4

Figure Lengend Snippet: Graphs for BAG2 immunostaining in RMP vs MPM regardless of subtype ( A ) and RMP vs the different subtypes of MPM ( B ), respectively. Graphs for MAD2L1 immunostaining in RMP vs MPM regardless of subtype ( C ) and RMP vs the different subtypes of MPM ( D ), respectively. Graphs for MDK immunostaining in RMP vs MPM regardless of subtype ( E ) and RMP vs the different subtypes of MPM ( F ), respectively. Kruskal–Wallis test followed by a post hoc Dunn’s test with Bonferroni correction was used to evaluate the statistical difference between the median of each group, p value <0.001(***), <0.01(**), <0.05(*).

Article Snippet: IHC was carried out on 3.5 μm paraffin sections by using recombinant Anti-BAG2 Rabbit mAb [EPR3567] (1:400), Anti-Midkine Rabbit mAb [EP1143Y] (1:100; cod ab79406, ab52637; Abcam, Cambridge, MA, USA), and Anti-MAD2 (MAD2L1) Rabbit pAb (1:250; cod TA308923; OriGene, Rockville, MD, USA).

Techniques: Immunostaining

Journal: Nature cell biology

Article Title: Ubiquitination-dependent localization of Polo-like kinase 1 in mitosis

doi: 10.1038/ncb2695

Figure Lengend Snippet: a, HeLa cells expressing H2B-mRFP were treated with the indicated siRNAs for 48 h and analyzed by live-cell microscopy. Stars indicate misaligned and lagging chromosomes. Bar is 10μm. b, The percentage of cells that complete mitosis normally or with a delay from prophase to anaphase (>50 min) or that undergo apoptosis during prometaphase was quantified (n = 50). c, The percentage of cells that undergo normal mitosis, become multinucleated or die during mitosis was quantified (n=50). d, HeLa cells were treated with the indicated siRNAs for 48 h and analyzed by immunofluorescence microscopy for MAD2 and kinetochores. The percentage of prometaphase and metaphase cells that contain one or more MAD2 positive kinetochores was quantified (n = 30). Example images are shown in Suppl. Fig. 1c. e, HeLa cells expressing H2B-mRFP were treated with the indicated mixtures of siRNAs for 48 h and analyzed by live-cell microscopy. The percentage of cells that die in prometaphase was quantified (n = 50). b-d, Bars represent the mean of three independent experiments. Error bars indicate +/-s.d.

Article Snippet: The following antibodies were used in the study: mouse monoclonal BubR1 (BD Biosciences 612502, 1:1000), pSer676 BubR1 (kind gift of E. A. Nigg, University of Basel, 1:???), CREST (Antibodies Incorporated, 15-234, 1:250), rabbit polyclonal CUL3 21 , mouse monoclonal FLAG (Sigma Aldrich F3165, 1:3000), mouse monoclonal GST (Labforce B-14, 1 :4000), rabbit polyclonal HA (Covance HA.11, 1 :3000), rabbit polyclonal KLHL22 and KLHL21 22 , rabbit polyclonal KLHL9 and 13 21 , rabbit polyclonal Mad2 (Bethyl Laboratories A300-301A, 1 :5000), mouse monoclonal MBP (Abcam ab49923, 1 :1000), rabbit polyclonal PLK1 (Abcam ab70697, 1:1000), mouse monoclonal PLK1 (Abcam ab17057, 1:500), rabbit polyclonal PLK2 (kind gift of I. Hoffmann, DKFZ Heidelberg, Germany), mouse monoclonal αTubulin (Sigma Aldrich T5169, IF 1:5000, WB 1:10000), rabbit polyclonal γTubulin (Sigma T3559, 1:1000), mouse monoclonal Cyclin A (Sigma C 4710, 1 :2000), mouse monoclonal Cyclin B1 (Santa Cruz GSN1, 1:5000), rabbit polyclonal phospho HistoneH3 (Upstate 06-570, 1:500), mouse monoclonal AuroraB (BD Biosciences 611082/3, 1:250), mouse monoclonal Hec1 (GeneTex, ??

Techniques: Expressing, Microscopy, Immunofluorescence