rabbit anti tpx2 (Bethyl)

Bethyl is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Bethyl rabbit anti tpx2

Identification of <t>TPX2/Aurora</t> A as 53BP1 effectors. (A) Flag-HA-53BP1 and associated proteins were isolated from HeLa-S nuclear extract using sequential Flag and HA immunoaffinity purification. The final eluted material was analyzed by SDS-PAGE and silver staining. (B) The 53BP1 complex from A was analyzed by MS/MS twice, and the total/unique peptide numbers for each protein are shown for each run. (C) Nuclear extracts from HeLa-S cells were immunoprecipitated with control IgG or <t>TPX2</t> antibody and Western blotted as shown. (D) GST, or the indicated GST-53BP1 fragments, were immobilized on glutathione-Sepharose, and binding with full-length recombinant His-TPX2 was tested. Bound material was analyzed by Western blot or Coomassie Blue staining. (E) Schematic of 53BP1 and summary of TPX2 binding data. (F) GST, GST-TPX2, or the indicated GST-TPX2 fragments were immobilized on glutathione-Sepharose and assessed for binding to full-length Flag-53BP1, as in D. (G) Schematic of TPX2 and summary of 53BP1 binding data. (H) GST, and increasing equimolar amounts of GST-TPX2 or GST- TPX2-Δ150-394, were tested for 53BP1 binding as done in F. (I) Recombinant His-Aurora A was immobilized on Ni-NTA and incubated with GST, GST-TPX2, or GST- TPX2-Δ150-394. Bound material was washed, subsequently incubated with Flag-53BP1, washed again, and analyzed for binding by Western blot or Coomassie Blue staining.

Rabbit Anti Tpx2, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+expression+430a+array+platform/TPX2+Antibody/pmc06363440-174-4-7
Average 93 stars, based on 13 article reviews

rabbit anti tpx2 - by Bioz Stars, 2026-11

93/100 stars

Images

1) Product Images from "Mitotic regulators TPX2 and Aurora A protect DNA forks during replication stress by counteracting 53BP1 function"

Article Title: Mitotic regulators TPX2 and Aurora A protect DNA forks during replication stress by counteracting 53BP1 function

Journal: The Journal of Cell Biology

doi: 10.1083/jcb.201803003

Identification of TPX2/Aurora A as 53BP1 effectors. (A) Flag-HA-53BP1 and associated proteins were isolated from HeLa-S nuclear extract using sequential Flag and HA immunoaffinity purification. The final eluted material was analyzed by SDS-PAGE and silver staining. (B) The 53BP1 complex from A was analyzed by MS/MS twice, and the total/unique peptide numbers for each protein are shown for each run. (C) Nuclear extracts from HeLa-S cells were immunoprecipitated with control IgG or TPX2 antibody and Western blotted as shown. (D) GST, or the indicated GST-53BP1 fragments, were immobilized on glutathione-Sepharose, and binding with full-length recombinant His-TPX2 was tested. Bound material was analyzed by Western blot or Coomassie Blue staining. (E) Schematic of 53BP1 and summary of TPX2 binding data. (F) GST, GST-TPX2, or the indicated GST-TPX2 fragments were immobilized on glutathione-Sepharose and assessed for binding to full-length Flag-53BP1, as in D. (G) Schematic of TPX2 and summary of 53BP1 binding data. (H) GST, and increasing equimolar amounts of GST-TPX2 or GST- TPX2-Δ150-394, were tested for 53BP1 binding as done in F. (I) Recombinant His-Aurora A was immobilized on Ni-NTA and incubated with GST, GST-TPX2, or GST- TPX2-Δ150-394. Bound material was washed, subsequently incubated with Flag-53BP1, washed again, and analyzed for binding by Western blot or Coomassie Blue staining.

Figure Legend Snippet: Identification of TPX2/Aurora A as 53BP1 effectors. (A) Flag-HA-53BP1 and associated proteins were isolated from HeLa-S nuclear extract using sequential Flag and HA immunoaffinity purification. The final eluted material was analyzed by SDS-PAGE and silver staining. (B) The 53BP1 complex from A was analyzed by MS/MS twice, and the total/unique peptide numbers for each protein are shown for each run. (C) Nuclear extracts from HeLa-S cells were immunoprecipitated with control IgG or TPX2 antibody and Western blotted as shown. (D) GST, or the indicated GST-53BP1 fragments, were immobilized on glutathione-Sepharose, and binding with full-length recombinant His-TPX2 was tested. Bound material was analyzed by Western blot or Coomassie Blue staining. (E) Schematic of 53BP1 and summary of TPX2 binding data. (F) GST, GST-TPX2, or the indicated GST-TPX2 fragments were immobilized on glutathione-Sepharose and assessed for binding to full-length Flag-53BP1, as in D. (G) Schematic of TPX2 and summary of 53BP1 binding data. (H) GST, and increasing equimolar amounts of GST-TPX2 or GST- TPX2-Δ150-394, were tested for 53BP1 binding as done in F. (I) Recombinant His-Aurora A was immobilized on Ni-NTA and incubated with GST, GST-TPX2, or GST- TPX2-Δ150-394. Bound material was washed, subsequently incubated with Flag-53BP1, washed again, and analyzed for binding by Western blot or Coomassie Blue staining.

Techniques Used: Isolation, Immunoaffinity Purification, SDS Page, Silver Staining, Tandem Mass Spectroscopy, Immunoprecipitation, Western Blot, Binding Assay, Recombinant, Staining, Incubation

TPX2 and Aurora A regulate BRCA1 recruitment and function. (A) U2OS cells expressing the indicated shRNAs were irradiated (5 Gy) and analyzed for 53BP1 and pH2A.X foci formation. Percentage of cells with ≥10 53BP1 foci ± SD is shown on the right. Scale bar, 10 µm. Foci were quantitated from three biological replicates with n ≥ 200 cells per experiment. Statistics: Student’s two-tailed t test; *, P < 0.05. (B) U2OS cells expressing the indicated shRNAs were treated with camptothecin (CPT) as shown, and chromatin-bound RPA was quantified by flow cytometry. Bar graph indicates the percentage of S/G2 cells that are RPA positive, as determined by 7-AAD staining from four biological replicates. Error bars represent ± SD. Statistics: Student’s two-tailed t test; *, P < 0.05. (C) Quantitation of HR efficiency of DR-GFP U2OS cells treated with the indicated shRNA and with or without the Flag-ISce expression vector. Cells were analyzed for GFP positivity via flow cytometry in three biological replicates as described in Materials and methods. Error bars indicate ± SD. Statistics: Student’s two-tailed t test; *, P < 0.05. (D and E) U2OS cells expressing the indicated shRNAs were irradiated (5 Gy) and analyzed for BRCA1 and p.H2A.X foci formation. Percentage of cells with ≥10 BRCA1 foci ± SD is shown on the right. Scale bar, 10 µm. Foci were quantitated from three biological replicates with n ≥ 250 cells per experiment. Statistics: Student’s two-tailed t test; *, P < 0.05. (F) Schematic of FUCCI cell cycle indicator expression pattern of mK02-Cdt (red; G1) and mAG-Geminin (green; S/G2/M; left). U2OS-FUCCI cells were treated with the indicated shRNAs, and S/G2-phase cells (yellow or green cells; indicated by white arrows) were analyzed for BRCA1 foci formation (right). Percentage of S/G2 cells with ≥10 BRCA1 foci ± SD is shown on the right. Scale bar, 10 µm. Foci were quantitated from three biological replicates with n ≥ 250 cells per experiment. Statistics: Student’s two-tailed t test; *, P < 0.05. (G and H) U2OS cells expressing the indicated shRNAs were irradiated (5 Gy) and analyzed for Rad51 foci. Percentage of cells with four or greater Rad51 foci ± SD is shown on the right. Scale bar, 10 µm. Foci were quantitated from three biological replicates with n ≥ 250 cells per experiment. Statistics: Student’s two-tailed t test; *, P < 0.05.

Figure Legend Snippet: TPX2 and Aurora A regulate BRCA1 recruitment and function. (A) U2OS cells expressing the indicated shRNAs were irradiated (5 Gy) and analyzed for 53BP1 and pH2A.X foci formation. Percentage of cells with ≥10 53BP1 foci ± SD is shown on the right. Scale bar, 10 µm. Foci were quantitated from three biological replicates with n ≥ 200 cells per experiment. Statistics: Student’s two-tailed t test; *, P < 0.05. (B) U2OS cells expressing the indicated shRNAs were treated with camptothecin (CPT) as shown, and chromatin-bound RPA was quantified by flow cytometry. Bar graph indicates the percentage of S/G2 cells that are RPA positive, as determined by 7-AAD staining from four biological replicates. Error bars represent ± SD. Statistics: Student’s two-tailed t test; *, P < 0.05. (C) Quantitation of HR efficiency of DR-GFP U2OS cells treated with the indicated shRNA and with or without the Flag-ISce expression vector. Cells were analyzed for GFP positivity via flow cytometry in three biological replicates as described in Materials and methods. Error bars indicate ± SD. Statistics: Student’s two-tailed t test; *, P < 0.05. (D and E) U2OS cells expressing the indicated shRNAs were irradiated (5 Gy) and analyzed for BRCA1 and p.H2A.X foci formation. Percentage of cells with ≥10 BRCA1 foci ± SD is shown on the right. Scale bar, 10 µm. Foci were quantitated from three biological replicates with n ≥ 250 cells per experiment. Statistics: Student’s two-tailed t test; *, P < 0.05. (F) Schematic of FUCCI cell cycle indicator expression pattern of mK02-Cdt (red; G1) and mAG-Geminin (green; S/G2/M; left). U2OS-FUCCI cells were treated with the indicated shRNAs, and S/G2-phase cells (yellow or green cells; indicated by white arrows) were analyzed for BRCA1 foci formation (right). Percentage of S/G2 cells with ≥10 BRCA1 foci ± SD is shown on the right. Scale bar, 10 µm. Foci were quantitated from three biological replicates with n ≥ 250 cells per experiment. Statistics: Student’s two-tailed t test; *, P < 0.05. (G and H) U2OS cells expressing the indicated shRNAs were irradiated (5 Gy) and analyzed for Rad51 foci. Percentage of cells with four or greater Rad51 foci ± SD is shown on the right. Scale bar, 10 µm. Foci were quantitated from three biological replicates with n ≥ 250 cells per experiment. Statistics: Student’s two-tailed t test; *, P < 0.05.

Techniques Used: Expressing, Irradiation, Two Tailed Test, Flow Cytometry, Staining, Quantitation Assay, shRNA, Plasmid Preparation

TPX2 and Aurora A promote replication fork stability. (A) Representative DNA fiber images of control, TPX2-depleted, and Aurora A–depleted U2OS cells treated with HU (4 mM) for 120 min. IdU, red; CldU, green. Scale bar, 50 µm. (B) Schematic of single-molecule DNA fiber track analysis (top) and size distribution of IdU track length in TPX2- and Aurora A–depleted cells in the presence and absence of HU or Mirin (bottom). Mirin (50 µM) was added concomitantly with HU treatment, as indicated. Out of two biological replicates; n ≥ 300 tracks were scored for each dataset. Bars represent median. Statistics: Mann–Whitney; ****, P < 0.0001; n.s., not significant. (C) Labeling and HU treatment schematic (top) and size distribution of IdU track length in U2OS cells expressing the indicated combination of shRNA and TPX2 expression vector as performed in B (bottom). (D) Size distribution of IdU track length in U2OS cells expressing the indicated combination of shRNA and Aurora A expression vector as performed in B. (E) U2OS cells were pulse labeled with 10 µM EdU and subjected to a 1-h thymidine chase (10 µM). Proteins bound to nascent DNA were purified by aniPOND as described in Materials and methods and analyzed by Western blot. (F) Size distribution of IdU track length in BRCA1-deficient (UWB1) and -proficient (UWB1 + BRCA1) cells in the presence of HU. Cells were infected with control or TPX2 shRNA before IdU and CldU labeling, as indicated. Fiber analysis was performed as in B.

Figure Legend Snippet: TPX2 and Aurora A promote replication fork stability. (A) Representative DNA fiber images of control, TPX2-depleted, and Aurora A–depleted U2OS cells treated with HU (4 mM) for 120 min. IdU, red; CldU, green. Scale bar, 50 µm. (B) Schematic of single-molecule DNA fiber track analysis (top) and size distribution of IdU track length in TPX2- and Aurora A–depleted cells in the presence and absence of HU or Mirin (bottom). Mirin (50 µM) was added concomitantly with HU treatment, as indicated. Out of two biological replicates; n ≥ 300 tracks were scored for each dataset. Bars represent median. Statistics: Mann–Whitney; ****, P < 0.0001; n.s., not significant. (C) Labeling and HU treatment schematic (top) and size distribution of IdU track length in U2OS cells expressing the indicated combination of shRNA and TPX2 expression vector as performed in B (bottom). (D) Size distribution of IdU track length in U2OS cells expressing the indicated combination of shRNA and Aurora A expression vector as performed in B. (E) U2OS cells were pulse labeled with 10 µM EdU and subjected to a 1-h thymidine chase (10 µM). Proteins bound to nascent DNA were purified by aniPOND as described in Materials and methods and analyzed by Western blot. (F) Size distribution of IdU track length in BRCA1-deficient (UWB1) and -proficient (UWB1 + BRCA1) cells in the presence of HU. Cells were infected with control or TPX2 shRNA before IdU and CldU labeling, as indicated. Fiber analysis was performed as in B.

Techniques Used: MANN-WHITNEY, Labeling, Expressing, shRNA, Plasmid Preparation, Purification, Western Blot, Infection

TPX2/Aurora A negatively regulate 53BP1 in the DNA damage response. (A) WT U2OS or 53BP1 knockout cells expressing the indicated shRNAs were irradiated (5 Gy) and analyzed for BRCA1 and pH2A.X foci formation. (B) U2OS cells treated as in A were also analyzed by Rad51 and pH2A.X foci. Scale bars, 10 µm. (C) Quantitation of BRCA1 foci from A. Foci were quantitated from three biological replicates with n ≥ 200 cells per experiment. Error bars indicate ±SD. Statistics: Student’s two-tailed t test; *, P < 0.05. (D) Quantitation of Rad51 foci from B. Foci were quantitated from three technical replicates with n ≥ 200 cells per experiment. Error bars indicate ±SD. Statistics: Student’s two-tailed t test; *, P < 0.05. (E) Size distribution of IdU track length in U2OS cells. WT, 53BP1 KO, or PTIP KO cells were treated with control or TPX2 shRNA before IdU and CldU labeling, as indicated. Out of two biological replicates; n ≥ 300 tracks scored for each dataset. Bars represent median. Statistics: Mann–Whitney; ****, P < 0.0001; n.s., not significant. (F) Size distribution of IdU track length in MEFs. WT or 53BP1 −/− cells were treated with control or TPX2 shRNA before labeling and HU treatment as indicated. Fiber analysis was performed as in E. (G) Size distribution of IdU track length in U2OS cells expressing the indicated combination of shRNA and TPX2 expression vector. Out of three biological replicates; n ≥ 300 tracks scored for each dataset. Bars represent median. Statistics: Mann–Whitney; ****, P < 0.0001; n.s., not significant. (H) U2OS WT or 53BP1-knockout cells were treated with the indicated shRNAs and subsequently treated with HU, as described in Materials and methods. Cell viability after 48 h of drug exposure was measured by MTS assay. Graph is a representative of three biological replicate experiments. Error bars indicate ±SD with n = 5 technical replicates.

Figure Legend Snippet: TPX2/Aurora A negatively regulate 53BP1 in the DNA damage response. (A) WT U2OS or 53BP1 knockout cells expressing the indicated shRNAs were irradiated (5 Gy) and analyzed for BRCA1 and pH2A.X foci formation. (B) U2OS cells treated as in A were also analyzed by Rad51 and pH2A.X foci. Scale bars, 10 µm. (C) Quantitation of BRCA1 foci from A. Foci were quantitated from three biological replicates with n ≥ 200 cells per experiment. Error bars indicate ±SD. Statistics: Student’s two-tailed t test; *, P < 0.05. (D) Quantitation of Rad51 foci from B. Foci were quantitated from three technical replicates with n ≥ 200 cells per experiment. Error bars indicate ±SD. Statistics: Student’s two-tailed t test; *, P < 0.05. (E) Size distribution of IdU track length in U2OS cells. WT, 53BP1 KO, or PTIP KO cells were treated with control or TPX2 shRNA before IdU and CldU labeling, as indicated. Out of two biological replicates; n ≥ 300 tracks scored for each dataset. Bars represent median. Statistics: Mann–Whitney; ****, P < 0.0001; n.s., not significant. (F) Size distribution of IdU track length in MEFs. WT or 53BP1 −/− cells were treated with control or TPX2 shRNA before labeling and HU treatment as indicated. Fiber analysis was performed as in E. (G) Size distribution of IdU track length in U2OS cells expressing the indicated combination of shRNA and TPX2 expression vector. Out of three biological replicates; n ≥ 300 tracks scored for each dataset. Bars represent median. Statistics: Mann–Whitney; ****, P < 0.0001; n.s., not significant. (H) U2OS WT or 53BP1-knockout cells were treated with the indicated shRNAs and subsequently treated with HU, as described in Materials and methods. Cell viability after 48 h of drug exposure was measured by MTS assay. Graph is a representative of three biological replicate experiments. Error bars indicate ±SD with n = 5 technical replicates.

Techniques Used: Knock-Out, Expressing, Irradiation, Quantitation Assay, Two Tailed Test, shRNA, Labeling, MANN-WHITNEY, Plasmid Preparation, MTS Assay

TPX2/Aurora A regulate DSB repair and replication fork stability via negative regulation of 53BP1. (A) Model of TPX2/Aurora A regulation of 53BP1 in response to double-strand breaks. (B) Model of TPX2/Aurora A function in response to replication stress. See text for details.

Figure Legend Snippet: TPX2/Aurora A regulate DSB repair and replication fork stability via negative regulation of 53BP1. (A) Model of TPX2/Aurora A regulation of 53BP1 in response to double-strand breaks. (B) Model of TPX2/Aurora A function in response to replication stress. See text for details.

Techniques Used:

Western Blot:

Article Title: Mitotic regulators TPX2 and Aurora A protect DNA forks during replication stress by counteracting 53BP1 function
Article Snippet: .. MS analysis of Flag-HA-53BP1 (Continued) Byrum et al. A fork protection pathway via TPX2/Aurora A and 53BP1 Journal of Cell Biology https://doi.org/10.1083/jcb.201803003 S21 Antibody Method Source Identifier Flag (mouse) WB and IP Sigma F3165 HA (mouse) WB, IF, and IP BioLegend 901501 β-Actin HRP (mouse) WB Sigma A3854 Alexa Fluor 488 (mouse) IF Invitrogen A11029 Alexa Fluor 488 (rabbit) IF Invitrogen A11034 Alexa Fluor 488 (rat) Flow cytometry and DNA fiber Invitrogen A11006 Alexa Fluor 594 (mouse) IF Invitrogen A11032 Alexa Fluor 594 (rabbit) IF Invitrogen A11037 Pacific Blue (mouse) IF (FUC CI) Invitrogen P31582 Aurora A (mouse) WB and IF Abcam ab13824 TPX2 (rabbit) WB Bethyl A300-430A TPX2 (rabbit) IF and IP Bethyl A300-429A 53BP1 (mouse) WB and IF BD Biosciences 612522 GAP DH WB Abcam ab181602 Histone H3 (rabbit) WB Abcam ab1791 GST (rabbit) WB Santa Cruz SC-459 Histone H3 pS10 (rabbit) WB Cell Signaling 9701S LSD1 (mouse) WB Santa Cruz SC-53875 BRCA1 (mouse) IF Santa Cruz SC-6954 BRCA1 (mouse) WB Abcam Ab16781 pH2AX (mouse) IF Abcam Ab26350 pH2AX (rabbit) IF Active Motif 39117 Rad51 (rabbit) IF Santa Cruz SC-8349 Rif1 (rabbit) IF Bethyl A300-569A RPA32 (rat) Flow cytometry Cell Signaling 2208S HRP (mouse) WB Cell Signaling 7076S HRP (rabbit) WB Cell Signaling 7074S BrdU (rat) DNA fiber Abcam ab6326 BrdU (mouse) DNA fiber BD Biosciences 347580 IF, immunofluorescence; IP, immunoprecipitation; WB, Western blot. ..

Article Title: Aurora kinase-A overexpression in mouse mammary epithelium induces mammary adenocarcinomas harboring genetic alterations shared with human breast cancer
Article Snippet: .. Western blot and antibodies For western blots, frozen tissues were lysed in radioimmunoprecipitation assay buffer containing protease inhibitors (Roche Diagnostics), resolved in sodium dodecyl sulfate–polyacrylamide gel electrophoresis, transferred and incubated with primary antibodies against human Aurora-A ( 4 ), Plk1, ERα, p53, Hsp90, β-actin (Santa Cruz Biotechnology), Tpx2 (Bethyl Laboratories), E-Cadherin (BD Biosciences), Her2/Neu, Cyclin D1, P-Akt (S473), P-Akt (T308), Akt and Vimentin (Cell Signaling). ..

Article Title: Aurora kinase-A overexpression in mouse mammary epithelium induces mammary adenocarcinomas harboring genetic alterations shared with human breast cancer
Article Snippet: .. For western blots, frozen tissues were lysed in radioimmunoprecipitation assay buffer containing protease inhibitors (Roche Diagnostics), resolved in sodium dodecyl sulfate–polyacrylamide gel electrophoresis, transferred and incubated with primary antibodies against human Aurora-A ( 4 ), Plk1, ERα, p53, Hsp90, β-actin (Santa Cruz Biotechnology), Tpx2 (Bethyl Laboratories), E-Cadherin (BD Biosciences), Her2/Neu, Cyclin D1, P-Akt (S473), P-Akt (T308), Akt and Vimentin (Cell Signaling). ..

Flow Cytometry:

Immunofluorescence:

Immunoprecipitation:

Article Title: Megakaryocytic Leukemia 1 Directs a Histone H3 Lysine 4 Methyltransferase Complex to Regulate Hypoxic Pulmonary Hypertension
Article Snippet: Vascular endothelium represents the first line of defense against stress stimuli guarding the integrity of the vasculature.. As such, disruption of endothelial homeostasis is intimately associated with a host of cardiovascular disorders, including atherosclerosis, cardiac infarction, and hypoxic pulmonary hypertension (HPH).. Endothelial injury in HPH is reflected by persistent vessel constriction, accumulation of reactive oxygen species, accelerated synthesis and deposition of extracellular matrix proteins, and build-up of proinflammatory milieu.

Staining:

Article Title: BRCA1 haploinsufficiency for replication stress suppression in primary cells.
Article Snippet: Cells were blocked with 5%BSA/PBST and then incubated with respective antibodies for 30min at 37 C followed by incubation with secondary antibodies (FITC or Rhodamine) for 30min at 37 C. Primary antibodies used in immunofluorescence studies were BRCA1 (Upstate; 1:500), phospho53BP1(S1778) (Cell Signaling, 2675S; 1:200), RPA (Cal Biochem, NA13; 1:100), 53BP1 (Bethyl Labs, A300-272A; 1:2,000), Rad51(Santa Cruz, SC-8349; 1:150), Mre11 (Genetex, GTX70212 1:200), CtIP (generous gift from Dr. Richard Baer), HA (Covance, MMS-101P; 1:500) and g-H2AX (Millipore, 05-636; 1:5,000). .. For TPX2 (Bethyl Labs, A300-429A; 1:400) and g-tubulin (Sigma- Aldrich, T6557; 1:1,000) staining, the cells were pre-fixed with acetone:methanol (3:7) at 20 C for 10min, followed by triton extraction (0.2% triton-X-100 in 20mM HEPES, pH 7.4, 50mM NaCl, 3mM MgCl2, 300mM Sucrose) at room temperature. ..

Extraction:

Incubation:

Article Title: Drug resistance dependent on allostery: A P-loop rigor Eg5 mutant exhibits resistance to allosteric inhibition by STLC
Article Snippet: .. Coverslips were placed in a humid chamber, incubated with a solution of primary antibodies: mouse monoclonal anti-tubulin clone TUB2.1 (Sigma: T4026) diluted at 1/450, human autoantibody against centromeres HCT-0100 ( Immunovision ) diluted at 1/450, mouse monoclonal anti-myc antibody (Covance, Berkeley, CA), rabbit polyclonal anti-TPX2 (Bethyl Laboratories, Motgomery, TX) diluted at 1/200 in antibody buffer (PBS 1X, sodium azide 0,02%, Tween 0,05%, and 3% (w/v) BSA) for 1 hour at 37°C. ..

Article Title: Aurora kinase-A overexpression in mouse mammary epithelium induces mammary adenocarcinomas harboring genetic alterations shared with human breast cancer
Article Snippet: .. For western blots, frozen tissues were lysed in radioimmunoprecipitation assay buffer containing protease inhibitors (Roche Diagnostics), resolved in sodium dodecyl sulfate–polyacrylamide gel electrophoresis, transferred and incubated with primary antibodies against human Aurora-A ( 4 ), Plk1, ERα, p53, Hsp90, β-actin (Santa Cruz Biotechnology), Tpx2 (Bethyl Laboratories), E-Cadherin (BD Biosciences), Her2/Neu, Cyclin D1, P-Akt (S473), P-Akt (T308), Akt and Vimentin (Cell Signaling). ..

Radio Immunoprecipitation:

Article Title: Aurora kinase-A overexpression in mouse mammary epithelium induces mammary adenocarcinomas harboring genetic alterations shared with human breast cancer
Article Snippet: .. For western blots, frozen tissues were lysed in radioimmunoprecipitation assay buffer containing protease inhibitors (Roche Diagnostics), resolved in sodium dodecyl sulfate–polyacrylamide gel electrophoresis, transferred and incubated with primary antibodies against human Aurora-A ( 4 ), Plk1, ERα, p53, Hsp90, β-actin (Santa Cruz Biotechnology), Tpx2 (Bethyl Laboratories), E-Cadherin (BD Biosciences), Her2/Neu, Cyclin D1, P-Akt (S473), P-Akt (T308), Akt and Vimentin (Cell Signaling). ..

Nucleic Acid Electrophoresis:

Article Title: Aurora kinase-A overexpression in mouse mammary epithelium induces mammary adenocarcinomas harboring genetic alterations shared with human breast cancer
Article Snippet: .. For western blots, frozen tissues were lysed in radioimmunoprecipitation assay buffer containing protease inhibitors (Roche Diagnostics), resolved in sodium dodecyl sulfate–polyacrylamide gel electrophoresis, transferred and incubated with primary antibodies against human Aurora-A ( 4 ), Plk1, ERα, p53, Hsp90, β-actin (Santa Cruz Biotechnology), Tpx2 (Bethyl Laboratories), E-Cadherin (BD Biosciences), Her2/Neu, Cyclin D1, P-Akt (S473), P-Akt (T308), Akt and Vimentin (Cell Signaling). ..

Review