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Bethyl anti mad2 rabbit polyclonal
Anti Mad2 Rabbit Polyclonal, supplied by Bethyl, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+mad2/pm39787108-98-51-54?v=Bethyl
Average 92 stars, based on 7 article reviews

anti mad2 rabbit polyclonal - by Bioz Stars, 2026-07

92/100 stars

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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.

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

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(****)).

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(****)).

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

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(****).

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(****).

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

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.

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.

Techniques: Expressing, Staining, Negative Staining

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(*).

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(*).

Techniques: Immunostaining

Fig. 3 A representative western blot of the siRNA transfection experiment. 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(****)).

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: Fig. 3 A representative western blot of the siRNA transfection experiment. 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 AntiBAG2 Rabbit mAb [EPR3567] (1:400), Anti-Midkine Rabbit mAb [EP1143Y] (1:100; cod ab79406, ab52637; Abcam, Cambridge, MA, USA), and AntiMAD2 (MAD2L1) Rabbit pAb (1:250; cod TA308923; OriGene, Rockville, MD, USA).

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

Fig. 4 Viability, proliferation and caspase assays after gene silencing. 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 (t0). 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(****).

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: Fig. 4 Viability, proliferation and caspase assays after gene silencing. 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 (t0). 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(****).

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

Fig. 7 Representative immunohistochemical staining of MPM samples showing intense expression of MDK, MAD2L1, and BAG2 proteins in tumor cells (brown staining). 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.

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: Fig. 7 Representative immunohistochemical staining of MPM samples showing intense expression of MDK, MAD2L1, and BAG2 proteins in tumor cells (brown staining). 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.

Techniques: Immunohistochemical staining, Staining, Expressing, Negative Staining

Fig. 8 Quantiﬁcation of protein expression in RMP vs MPM as assessed by IHC and H-score. 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(*).

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: Fig. 8 Quantiﬁcation of protein expression in RMP vs MPM as assessed by IHC and H-score. 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(*).

Techniques: Expressing, Immunostaining

Cdk1 phoshorylation of Cdc20 is required for an efficient mitotic arrest in response to microtubule poisons. (A) Live cell imaging of Hela cells depleted of endogenous Cdc20 and transiently expressing untagged Cdc20 WT, 3DD, or 3AA in the presence of nocadazole (30 ng/ml). Percentage of cells that exit mitosis during the course of imaging from three independent experiments (mean and SD indicated). (B) As in A, time from nuclear envelope breakdown (NEBD) to mitotic exit or end of filming from three independent experiments (each dot represents a single cell, red line indicates mean and SD, n values are shown in the graph and represent single cells analyzed per condition; statistical analysis with Mann-Whitney test; ****P > 0.0001). (C) Live cell imaging of stable cell lines that were depleted of endogenous Cdc20 and expressing indicated YFP-tagged Cdc20 proteins in the presence of nocodazole (30 ng/ml). The R132A mutation prevents Mad2 binding, while the 4A mutations prevent BubR1 binding. Time from NEBD to mitotic exit (each dot represents a single cell, red line indicates mean and standard deviation, n values are shown in the graph and represent single cells analyzed per condition, statistical analysis with Mann-Whitney test; ****P > 0.0001). (D) Model: MCC-bound BubR1 is preferentially dephosphorylated by PP2A-B56, whereas free Cdc20 is phosphorylated by Cdk1.

Journal: The Journal of Cell Biology

Article Title: Coupling of Cdc20 inhibition and activation by BubR1

doi: 10.1083/jcb.202012081

Figure Lengend Snippet: Cdk1 phoshorylation of Cdc20 is required for an efficient mitotic arrest in response to microtubule poisons. (A) Live cell imaging of Hela cells depleted of endogenous Cdc20 and transiently expressing untagged Cdc20 WT, 3DD, or 3AA in the presence of nocadazole (30 ng/ml). Percentage of cells that exit mitosis during the course of imaging from three independent experiments (mean and SD indicated). (B) As in A, time from nuclear envelope breakdown (NEBD) to mitotic exit or end of filming from three independent experiments (each dot represents a single cell, red line indicates mean and SD, n values are shown in the graph and represent single cells analyzed per condition; statistical analysis with Mann-Whitney test; ****P > 0.0001). (C) Live cell imaging of stable cell lines that were depleted of endogenous Cdc20 and expressing indicated YFP-tagged Cdc20 proteins in the presence of nocodazole (30 ng/ml). The R132A mutation prevents Mad2 binding, while the 4A mutations prevent BubR1 binding. Time from NEBD to mitotic exit (each dot represents a single cell, red line indicates mean and standard deviation, n values are shown in the graph and represent single cells analyzed per condition, statistical analysis with Mann-Whitney test; ****P > 0.0001). (D) Model: MCC-bound BubR1 is preferentially dephosphorylated by PP2A-B56, whereas free Cdc20 is phosphorylated by Cdk1.

Article Snippet: The following antibodies were used at the indicated dilutions for Western blot: Cdc20 mouse monoclonal antibody (1:1,000; sc-13162; Santa Cruz Biotechnology; clone E7, indicated as #1), Cdc20 mouse monoclonal antibody (MAB3775; Millipore; clone AR12, indicated as #2), Cdc20 T70 and T59 phosphorylation-specific antibodies rabbit polyclonal antibody [raised against peptides CSKVQT(Tp)PSKPG and CRTPGR(Tp)PGKSS; Moravian Biotechnology; reference 22], APC3 T447p rabbit polyclonal [raised against Peptides CGKISTI(Tp)PQIQAF; Moravian Biotechnology], anti-phospho-histone H3 (Ser 10) rabbit polyclonal antibody (1:1,000; 06–570; Millipore), APC4 mouse monoclonal antibody (1:500; Moravian Biotechnology; clone CIV 1.1), APC3 mouse monoclonal antibody (1:500; Moravian Biotechnology), Mad2 rabbit polyclonal antibody (1:500; A300-300A; Bethyl Laboratories), Bub3 mouse monoclonal antibody (1:500; 611731; BD Biosciences), BubR1 rabbit polyclonal antibody (1:1,000; A300-995A; Bethyl Laboratories), and BubR1 mouse monoclonal antibody (1:500; Biotech Research and Innovation Centers; raised against TPR domain).

Techniques: Live Cell Imaging, Expressing, Imaging, MANN-WHITNEY, Stable Transfection, Mutagenesis, Binding Assay, Standard Deviation

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.

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

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