Journal: PLoS ONE

Article Title: Inhibitory Effect of Tumor Suppressor p53 on Proinflammatory Chemokine Expression in Ovarian Cancer Cells by Reducing Proteasomal Degradation of IκB

doi: 10.1371/journal.pone.0051116

Figure Lengend Snippet: (A) Signature of chemokine ligands and (B) chemokine receptors in human ovarian cancer cell lines. After isolating total RNA from each cell line, PCR array was performed using a customized PCR array plate containing complementary sequences for human chemokine genes. Different colors indicate average cycle threshold with expression ranges from >35 to <25. (C) Protein expression of p53 and Mdm2 in ovarian cancer cell lines. Whole cell lysates were prepared and Western blot was carried out using antibodies specific to p53, Mdm2 and β-actin as loading control. Experiments were performed in duplicate and a representative result is shown. OV, OVCAR-3 cells; SK, SKOV-3 cells; A, A2780 cells; Ca, CaOV-3 cells; TOV, TOV-21G cells.

Article Snippet: Recombinant human TNF and human p53 DuoSet® IC ELISA kit were obtained from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Western Blot, Control

Journal: PLoS ONE

Article Title: Inhibitory Effect of Tumor Suppressor p53 on Proinflammatory Chemokine Expression in Ovarian Cancer Cells by Reducing Proteasomal Degradation of IκB

doi: 10.1371/journal.pone.0051116

Figure Lengend Snippet: (A) TNF-induced chemokines in SKOV-3 cells. After isolating total RNA, PCR array was performed using a human chemokine PCR array plate. Dotted line indicates 2-fold increase; chemokines with a greater than 2-fold increase are recognized as TNF-induced chemokines. (B) Confirmation of p53 protein expression after transient transfection in SKOV-3 cells. After transfection of empty vector (EM) and p53 expression vector (p53), whole cell lysates were prepared and p53 expression was confirmed by Western blot. β-actin is used as a loading control. (C) Effect of p53 on TNF-induced chemokines. After overnight transfection of vectors, cells were treated with TNF (10 ng/ml) for 1 h and qRT-PCR was carried out using primers for CCL2, CXCL1, 2, 3 and 8. β-actin serves as normalization control. Different letters indicate significant differences (P≤0.05) within each chemokine group (ANOVA and Tukey's pairwise comparisons). Experiments were performed in triplicate and all data are shown as mean ± SE.

Article Snippet: Recombinant human TNF and human p53 DuoSet® IC ELISA kit were obtained from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot, Control, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: Inhibitory Effect of Tumor Suppressor p53 on Proinflammatory Chemokine Expression in Ovarian Cancer Cells by Reducing Proteasomal Degradation of IκB

doi: 10.1371/journal.pone.0051116

Figure Lengend Snippet: (A) Nucleotide sequences of promoters for TNF-induced chemokines such as CCL20, CXCL1, 2, 3 and 8. These chemokine promoters contain one NF-κB site at the proximal region, except for CCL20, which has two NF-κB sites at the distal and proximal region. (B) Effect of p53 on NF-κB luciferase activity. After transfection of vectors or cotransfection with p65, cells were treated with TNF (10 ng/ml) for 4 h. (C) Effect of p53 on TNF-activated IκB. After transfection of empty vector or p53 in SKOV-3 (p53 null), OVCAR-3 (p53 mutant) and A2780 (p53 wild-type), cells were treated with TNF (10 ng/ml) for indicated times. β-actin serves as loading control. Experiments were performed in duplicate and a representative result is shown; numbers below are relative density values.

Article Snippet: Recombinant human TNF and human p53 DuoSet® IC ELISA kit were obtained from R&D Systems (Minneapolis, MN).

Techniques: Luciferase, Activity Assay, Transfection, Cotransfection, Plasmid Preparation, Mutagenesis, Control

Journal: PLoS ONE

Article Title: Inhibitory Effect of Tumor Suppressor p53 on Proinflammatory Chemokine Expression in Ovarian Cancer Cells by Reducing Proteasomal Degradation of IκB

doi: 10.1371/journal.pone.0051116

Figure Lengend Snippet: (A) Accumulated effect of p53 on ubiquitylated proteins. After transient transfection of p53 in A2780, OVCAR-3 and SKOV-3 cells, whole cell lysates were prepared and Western blot was carried out using antibodies specific to ubiquitin, p21, IκB, p53 and β-actin (as loading control). Experiments were performed in duplicate and a representative result is shown. (B) Confirmation of p53 activity after transient transfection of p53. ELISA was performed in triplicate and data are shown as mean ± SE. Dark gray bars indicate significance (p<0.05, paired Student's t -test) within each cell line. (C) The effect of p53 on proteasome activity. Assays were performed in triplicate and data are shown as mean ± SE. Dark gray bars indicate significance (p<0.05, paired Student's t -test) within each cell line. (D) Effects of p53 on ubiquitination of IκB. Immunoprecipitated IκB was immunoblotted using ubiquitin antibody. Experiments were performed in duplicate and a representative result is shown.

Article Snippet: Recombinant human TNF and human p53 DuoSet® IC ELISA kit were obtained from R&D Systems (Minneapolis, MN).

Techniques: Transfection, Western Blot, Ubiquitin Proteomics, Control, Activity Assay, Enzyme-linked Immunosorbent Assay, Immunoprecipitation

Journal: PLoS ONE

Article Title: Inhibitory Effect of Tumor Suppressor p53 on Proinflammatory Chemokine Expression in Ovarian Cancer Cells by Reducing Proteasomal Degradation of IκB

doi: 10.1371/journal.pone.0051116

Figure Lengend Snippet: Effect of p53 on ubiquitin-activating enzymes (E1), ubiquitin-conjugating enzymes (E2) and ubiquitin-protein ligases (E3) obtained from comparison between empty vector and p53 vector transfected ovarian cancer cells.

Article Snippet: Recombinant human TNF and human p53 DuoSet® IC ELISA kit were obtained from R&D Systems (Minneapolis, MN).

Techniques: Ubiquitin Proteomics, Comparison, Plasmid Preparation, Transfection

Journal: PLoS ONE

Article Title: Inhibitory Effect of Tumor Suppressor p53 on Proinflammatory Chemokine Expression in Ovarian Cancer Cells by Reducing Proteasomal Degradation of IκB

doi: 10.1371/journal.pone.0051116

Figure Lengend Snippet: (A) Effect of p53 on Mdm2 expression. After transient transfection of p53 in A2780, OVCAR-3 and SKOV-3 cells, whole cell lysates were prepared and Western blot was carried out using antibodies specific to Mdm2; and β-actin served as loading control. (B) Effects of p53 expression on p53 binding to p65 and IκB. After transient transfection of p53, immunoprecipitated (IP) p53 was immunoblotted (IB) using p65 or IκB antibody. (C) Effect of p53 on expression of various IKK isoforms. After transient transfection of p53, whole cell lysates were prepared and Western blot was carried out using antibodies specific to IKKα, IKKβ, IKKγ, IKKε; β-actin served as loading control. Experiments were performed in duplicate and a representative result is shown.

Article Snippet: Recombinant human TNF and human p53 DuoSet® IC ELISA kit were obtained from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Transfection, Western Blot, Control, Binding Assay, Immunoprecipitation

Journal: PLoS ONE

Article Title: Inhibitory Effect of Tumor Suppressor p53 on Proinflammatory Chemokine Expression in Ovarian Cancer Cells by Reducing Proteasomal Degradation of IκB

doi: 10.1371/journal.pone.0051116

Figure Lengend Snippet: (A) Effect of nutlin-3 on NF-κB luciferase activity. After transfection of vectors or cotransfection with p53, cells were pretreated with nutlin-3 (10 µM) for 24 h followed by TNF (10 ng/ml) for 4 h. Different letters indicate significant differences (P≤0.05) within each group (ANOVA and Tukey's pairwise comparisons). Experiments were performed in triplicate and all data are shown as mean ± SE. (B) Effect of nutlin-3 on TNF-activated IκB. After transfection of empty vector or p53 in SKOV-3 cells, cells were pretreated with nutlin-3 (10 µM) for 24 h followed by TNF (10 ng/ml) for indicated times. β-actin serves as loading control. Experiments were performed in duplicate and a representative result is shown. (C) Effect of nutlin-3 on TNF-induced chemokines. After overnight transfection of vectors, cells were pretreated with nutlin-3 (10 µM) for 24 h followed by TNF (10 ng/ml) for 1 h and qRT-PCR was carried out using primers for CCL2, CXCL1, 2, 3 and 8. β-actin serves as normalization control. Asterisk indicates significant differences (P≤0.05, paired Student's t -test) when compared to the presence of nutlin-3. Experiments were performed in triplicate and all data are shown as mean ± SE.

Article Snippet: Recombinant human TNF and human p53 DuoSet® IC ELISA kit were obtained from R&D Systems (Minneapolis, MN).

Techniques: Luciferase, Activity Assay, Transfection, Cotransfection, Plasmid Preparation, Control, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: Inhibitory Effect of Tumor Suppressor p53 on Proinflammatory Chemokine Expression in Ovarian Cancer Cells by Reducing Proteasomal Degradation of IκB

doi: 10.1371/journal.pone.0051116

Figure Lengend Snippet: Chronic inflammation promotes ovarian cancer progression via NF-κB signaling. Wild-type p53 reduces activity of the ubiquitin-proteasome system, resulting in low IκB degradation (blue line). This reduces NF-κB activity, inhibiting proinflammatory chemokine expression and attenuating the proinflammatory tumor microenvironment (blue arrow). On the other hand, p53 increases Mdm2 expression (dark arrow) in a feedback loop to compensate for the reduced activity of the ubiquitin-proteasome system. Loss of p53 observed frequently in advanced ovarian cancer triggers high proinflammatory chemokines by increasing NF-κB signaling which is composed of IκB and p65/p50 followed by a high IκB degradation (red arrow). Enhanced NF-κB activity results in potentiation of the proinflammatory tumor microenvironment for ovarian cancer progression such as peritoneal tumor dissemination and massive ascites. The imbalance between p53 and Mdm2 also contributes to increasing NF-κB signaling via the ubiquitin-proteasome system.

Article Snippet: Recombinant human TNF and human p53 DuoSet® IC ELISA kit were obtained from R&D Systems (Minneapolis, MN).

Techniques: Activity Assay, Ubiquitin Proteomics, Expressing

Journal: Alcohol (Fayetteville, N.Y.)

Article Title: Profiling the Oxylipidome in Aged Mice after Chronic Ethanol Feeding: Identifying Lipid Metabolites as Drivers of Hepatocyte Stress

doi: 10.1016/j.alcohol.2022.08.012

Figure Lengend Snippet: Young and aged adult WT mice were allowed free access to ethanol (32%,d25) or pair-fed control diets. A) Liver RNA was isolated and expression of senescence markers, p16 and p21 mRNA was detected in mouse livers using qRT-PCR. B/C) Paraffin-embedded liver sections were deparaffinized followed by immunodetection of PCNA and p53 and nuclei were counterstained with hematoxylin. Frozen liver sections were stained for β-galactosidase activity. Images were acquired using a ×10 objective and positive staining was quantified using Image-J. Data are from two independent trials; individual values are reported and represented as means ± SEM, n=8 pair-fed and 12 EtOH-fed mice/group. *Values were significantly different from each other (P<0.05).

Article Snippet: Paraffin-embedded liver sections were deparaffinized and stained with antibodies against NIMP-R14 (Novus Biologicals, cat NB600–1387), PCNA (Millipore, cat# MAB424R) and p53 (Novus Biologicals, cat# NB200–103).

Techniques: Control, Isolation, Expressing, Quantitative RT-PCR, Immunodetection, Staining, Activity Assay

Journal: Nature chemical biology

Article Title: Targeting a Helix-in-Groove Interaction Between E1 and E2 Blocks Ubiquitin Transfer

doi: 10.1038/s41589-020-0625-7

Figure Lengend Snippet: (a) Dose-responsive inhibition of UBE1 by SAH-UBE2A in the presence and absence of 0.01% Triton X-100, as monitored by thioester transfer assay, gel electrophoresis, and silver stain. (b) Dose-responsive inhibition of UBE1 by SAH-UBE2A using reaction buffer containing 50 mM (standard; ) or 250 mM (elevated) NaCl, as monitored by thioester transfer assay, gel electrophoresis, and silver stain. (c) Fluorescent bead binding assay showing that FITC-SAH-UBE2A exhibits no non-specific binding to Ni-NTA beads (top row), selective binding to His-UBE1 coated beads (middle row), and no non-specific binding to beads coated with a negative control protein, such as His-p53 (bottom row). Scale bars, 200 μm. (d) VLCAD enzymatic assay performed in the presence of vehicle or SAH-UBE2A (30:1 peptide:enzyme ratio, final peptide concentration 43.2 μM), demonstrating no interference by SAH-UBE2A on the capacity of VLCAD to oxidize its palmitoyl-CoA substrate. Data are mean ± s.e.m. for experiments performed in technical triplicate. (e) In vitro ubiquitin pathway reconstitution assay showing that SAH-UBE2A (30 μM, excess) inhibits ubiquitination of p53 when the cascade relies on E1 thioester transfer activity but not when the E1 step is bypassed by the addition of E2 pre-charged with ubiquitin. In addition, SAH-UBE2A has no effect on the capacity of the DUB USP7 to deubiquitinate p53. The entire experiment was performed twice using independent preparations of reagents with similar results. (f) SAH-UBE2A has no effect on the capacity of UBE1 to form a covalent UBE1~ubiquitin thioester adduct, the enzymatic step of the UBE1 catalytic cycle that precedes thioester transfer to E2, as demonstrated by the presence of a UBE1 and UBE1~Ub doublet (silver stain) across all SAH-UBE2A concentrations of the thioester transfer assay. (g) To verify that the E1-inhibitory mechanism was not dependent on ubiquitin discharge from E2 onto SAH-UBE2A, a lysine-free K14R mutant was synthesized and tested in the thioester transfer assay. Both SAH-UBE2A and SAH-UBE2A K14R (10 μM dosing) showed equivalent E1-inhibitory activity, as reflected by blockade of E2~Ub formation. Uncropped gels for panels a, b, e-f are available online. Source data for the VLCAD enzymatic assay plot are available online.

Article Snippet: The membranes were blocked with 3% milk for 1 h, incubated overnight in PBS containing 3% BSA and mouse p53 antibody (Boston Biochem K-200B) at a 1:1000 dilution, washed in PBS containing 0.1% Tween-20, and then incubated in PBS containing 3% BSA and anti-mouse secondary antibody (BioRad AAC10P) at a 1:5000 dilution for 1 hour at room temperature.

Techniques: Inhibition, Nucleic Acid Electrophoresis, Silver Staining, Binding Assay, Negative Control, Enzymatic Assay, Concentration Assay, In Vitro, Ubiquitin Proteomics, Reconstitution Assay, Activity Assay, Mutagenesis, Synthesized

Journal: Cell Death and Differentiation

Article Title: FAS apoptotic inhibitory molecule 2 is a stress-induced intrinsic neuroprotective factor in the retina

doi: 10.1038/cdd.2017.109

Figure Lengend Snippet: FAIM2 interacts with p53 and HSP90 after FAS activation. (a,b) Immunoprecipitation of protein lysates from 661 W cells treated with FAS-ligand for 24 h with FAIM2 followed by liquid chromatography–tandem mass spectrometry (LC–MS/MS) proteomic analysis of interactors. The peptide abundance of p53 and HSP90 was quantified using normalized spectral abundance factor (NSAF) analysis. Data is representative of two independent experiments. Significant increase of p53 and HSP90 is observed in the FAIM2 complex of proteins immunoprecipitated after FAS receptor activation. (c) 661 W cells were treated with 500 ng/ml of FAS-ligand and incubated for 24 h. Proteins from whole cell lysate were immunoprecipitated with Faim2 antibody and immunoblotted with p53 antibody. Protein lysates were also immunoprecipitated with p53 antibody and immunoblotted with FAIM2 antibody to demonstrate FAIM2/p53 interaction, N=3. (d) Protein from 661 W cells treated with FAS-ligand for 24 h were immunoprecipitated with FAIM2. HSP90 antibody was used for Western blotting, showing an increase in HSP90/FAIM2 interaction after FAS receptor activation. Incubation with λ-phosphatase showed decreased levels of HSP90/FAIM2 binding, N=3. (e) Protein lysates from 661 W cells treated with FAS-ligand for 24 h with vehicle or 10 μM of HSP90 inhibitor geldanamycin (GD) were analyzed for FAIM2 levels. Inhibition of HSP90 activity with GD led to decreased FAIM2 levels in cells following FAS receptor activation, N=3

Article Snippet: Antibodies LFG (Cat. #: AS-54488, Anaspec, Fremont, CA, USA), Phosphothreonine (Cat. #: AB1607, EMD Millipore, Billerica, MA, USA), Phosphoserine (Cat. #: AB1603, EMD Millipore), p53 (Cat. #: AF1355, R&D Systems, Minneapolis, MN, USA), Phospho-HSP90 α (Thr5/7) (Cat. #3488, Cell Signaling Technology), HSP90 (Cat.#4874, Cell Signaling Technology), β -Actin (A5316-100UL, Sigma-Aldrich, St. Louis, MO, USA), Ubiquitin PD-40 (Cat.#, Cell Signaling Technology).

Techniques: Activation Assay, Immunoprecipitation, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Incubation, Western Blot, Binding Assay, Inhibition, Activity Assay

Journal: Food & Nutrition Research

Article Title: Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

doi: 10.29219/fnr.v65.5649

Figure Lengend Snippet: Primer sequences used in this study.

Article Snippet: Recombinant human p53 protein was obtained from Boston Biochem (Beijing, China).

Techniques:

Journal: Food & Nutrition Research

Article Title: Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

doi: 10.29219/fnr.v65.5649

Figure Lengend Snippet: Molecular docking and dynamics and of NOB and p53 protein. (a) The time dependence of root mean square deviation (RMSD) between NOB and p53 protein by molecular dynamic. (b) The time evolution of the radius of gyration (Rg) between NOB and p53 protein by molecular dynamic. (c) The molecular docking of NOB and p53 protein.

Article Snippet: Recombinant human p53 protein was obtained from Boston Biochem (Beijing, China).

Techniques:

Journal: Food & Nutrition Research

Article Title: Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

doi: 10.29219/fnr.v65.5649

Figure Lengend Snippet: Docking energies of NOB with p53 protein

Article Snippet: Recombinant human p53 protein was obtained from Boston Biochem (Beijing, China).

Techniques: Binding Assay

Journal: Food & Nutrition Research

Article Title: Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

doi: 10.29219/fnr.v65.5649

Figure Lengend Snippet: The interaction between NOB and p53 protein was verified in vitro . (a) UV-visible spectrum of the interaction between NOB and p53 protein. (b) Fluorescence spectrum of the interaction between NOB and p53 protein. Experimental conditions: 37°C, p53 protein was mixed with the same volume of NOB and maintained for 10 min. The final concentration of p53 was 1 μM, and those of NOB were 0, 10, 33, or 100 μM.

Article Snippet: Recombinant human p53 protein was obtained from Boston Biochem (Beijing, China).

Techniques: In Vitro, Fluorescence, Concentration Assay

Journal: Food & Nutrition Research

Article Title: Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

doi: 10.29219/fnr.v65.5649

Figure Lengend Snippet: The binding constant of NOB and p53 protein

Article Snippet: Recombinant human p53 protein was obtained from Boston Biochem (Beijing, China).

Techniques: Binding Assay

Journal: Food & Nutrition Research

Article Title: Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

doi: 10.29219/fnr.v65.5649

Figure Lengend Snippet: Secondary structure of p53 protein adding various NOB

Article Snippet: Recombinant human p53 protein was obtained from Boston Biochem (Beijing, China).

Techniques:

Journal: Food & Nutrition Research

Article Title: Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

doi: 10.29219/fnr.v65.5649

Figure Lengend Snippet: The effect of NOB on expressions of p53 pathway genes of hypoxia-induced JEG-3 and BeWo cells. The mRNA expressions of p53, MDM2, p21, BCL2, and BAX were estimated by RT-qPCR. The mRNA ratio of BCL2/BAX was calculated. (a) The mRNA expression analysis of JEG-3 cells. (b) The mRNA expression analysis of BeWo cells. The JEG-3 and BeWo cells were, respectively, exposed to 0, 10, 33, or 100 μM NOB with 500 μM cobalt chloride in Serum-free medium for 12 h. Cells treated without cobalt chloride and NOB for 12 h were taken as the normoxic control group. The values are presented as the mean ± SD of independent experiments ( n = 3). All different capital letters indicate significant differences at P < 0.05 using one-way ANOVA.

Article Snippet: Recombinant human p53 protein was obtained from Boston Biochem (Beijing, China).

Techniques: Quantitative RT-PCR, Expressing, Control

Journal: Food & Nutrition Research

Article Title: Nobiletin, a hexamethoxyflavonoid from citrus pomace, attenuates G1 cell cycle arrest and apoptosis in hypoxia-induced human trophoblast cells of JEG-3 and BeWo via regulating the p53 signaling pathway

doi: 10.29219/fnr.v65.5649

Figure Lengend Snippet: The effect of NOB on expressions of p53 pathway proteins of hypoxia-induced JEG-3 and BeWo cells. The protein expressions of p53, MDM2, p21, BCL2 and BAX were estimated by Western blot analysis. The protein ratio of BCL2/BAX was calculated. (a) The Western blot of JEG-3 cells. (b) The Western blot of BeWo cells. (c) The protein expression analysis of JEG-3 cells. (d) The protein expression analysis of BeWo cells. The JEG-3 and BeWo cells were, respectively, exposed to 0, 10, 33, or 100 μM NOB with 500 μM cobalt chloride in Serum-free medium for 12 h. Cells treated without cobalt chloride and NOB for 12 h were taken as the normoxic control group. The values are presented as the mean ± SD of independent experiments ( n = 3). All different capital letters indicate significant differences at P < 0.05 using one-way ANOVA.

Article Snippet: Recombinant human p53 protein was obtained from Boston Biochem (Beijing, China).

Techniques: Western Blot, Expressing, Control

Journal: Analytical and bioanalytical chemistry

Article Title: Anti-heat shock protein autoantibody profiling in breast cancer using customized protein microarray.

doi: 10.1007/s00216-015-9257-2

Figure Lengend Snippet: Fig. 2 Scheme of protein chip design. Each micro-well contains streptavidin-F555, buffer, HSPB1, HSPD1, HSP70, p53, HSP90, HSPA5, HSP90B1, and HSP110 spotted in five replicates. Lines 1 and 2 were incubated with purified antibody for positive control, lines 3 to 6 were incu- bated with sera from breast cancer patients, and lines 7–10 were incubated with healthy donor sera

Article Snippet: HSPB1 (adi-esp-715-d), HSP70 (adi-nsp-555-d), HSP90 (adi-spp-776-d), mouse-anti-human anti-HSPB1 antibody-biotin (adi-spa-800B-F), mouse-anti-human anti-HSP70 antibody-biotin (adi-spa-810B-F), and mouse-anti-human antiHSP90 antibody (adi-spa-831-200) were all purchased from Enzo Life Sciences (Switzerland); HSPD1 (ab113177), HSPA5 (ab78432), HSP90B1 (ab188463), HSP110 (ab78790), rabbit-anti-human anti-HSPD1 antibody-biotin (ab105853), and mouse-anti-human anti-HSP90B1 antibody (ab63469) were obtained fromAbcam (UK); p53 (p6249) and mouse-anti-human anti-p53 antibody-biotin (MA5-12554) were obtained from Sigma and Thermo Scientific (USA), respectively; mouse-anti-human anti-HSPA5 antibody (MAB4846) and mouse-anti-human anti-HSP110 antibody (MAB4029) were obtained from R&D Systems (USA); F555-labeled streptavidin (S-21381) was purchased from Invitrogen; and Cy3-labeled goat anti-human antibody immunoglobulin G (IgG) (109-165-008) and Cy3-labeled goat antimouse antibody IgG (115-165-008) were purchased from Jackson Immuno Research (USA).

Techniques: Incubation, Purification, Positive Control

Journal: Analytical and bioanalytical chemistry

Article Title: Anti-heat shock protein autoantibody profiling in breast cancer using customized protein microarray.

doi: 10.1007/s00216-015-9257-2

Figure Lengend Snippet: Fig. 6 Receiver operating characteristic (ROC) curve analysis of seven individual autoantibodies and of the combination of these autoantibodies to discriminate breast cancer patients from healthy controls. The detection of autoantibodies against HSPD1, HSP70, HSP90, and HSP90B1 was obtained on COOH surface; the detection of autoantibodies against HSPB1, P53, and HSPA5 was obtained on chitosan surface

Article Snippet: HSPB1 (adi-esp-715-d), HSP70 (adi-nsp-555-d), HSP90 (adi-spp-776-d), mouse-anti-human anti-HSPB1 antibody-biotin (adi-spa-800B-F), mouse-anti-human anti-HSP70 antibody-biotin (adi-spa-810B-F), and mouse-anti-human antiHSP90 antibody (adi-spa-831-200) were all purchased from Enzo Life Sciences (Switzerland); HSPD1 (ab113177), HSPA5 (ab78432), HSP90B1 (ab188463), HSP110 (ab78790), rabbit-anti-human anti-HSPD1 antibody-biotin (ab105853), and mouse-anti-human anti-HSP90B1 antibody (ab63469) were obtained fromAbcam (UK); p53 (p6249) and mouse-anti-human anti-p53 antibody-biotin (MA5-12554) were obtained from Sigma and Thermo Scientific (USA), respectively; mouse-anti-human anti-HSPA5 antibody (MAB4846) and mouse-anti-human anti-HSP110 antibody (MAB4029) were obtained from R&D Systems (USA); F555-labeled streptavidin (S-21381) was purchased from Invitrogen; and Cy3-labeled goat anti-human antibody immunoglobulin G (IgG) (109-165-008) and Cy3-labeled goat antimouse antibody IgG (115-165-008) were purchased from Jackson Immuno Research (USA).

Techniques: