Journal: Archives of Medical Science : AMS

Article Title: The decrease in hippocampal transient receptor potential M2 (TRPM2) channel and muscarinic acetylcholine receptor 1 (CHRM1) is associated with memory loss in a surgical menopause rat model

doi: 10.5114/aoms.2019.83760

Figure Lengend Snippet: TRPM2 and CHRM1 gene expression levels were lower in G2 compared with G1 a Compared with G1 (p < 0.05).

Article Snippet: In order to prevent surface staining, after treatment with Ultra V Block (TA-125-UB, the Lab Vision Corporation, USA) solutions, the tissues were incubated with primary antibodies for 60 min (CHRM1 was purchased from Boster (Cholinergic receptor, muscarinic 1, catalog number: PA2202, Boster, 3942 B Valley Ave, Pleasanton, CA, 94566) and TRPM2 was purchased from Abcam (Rabbit Anti-TRPM2 antibody, ab101738, Abcam, Cambridge, UK)).

Techniques: Gene Expression

Journal: Archives of Medical Science : AMS

Article Title: The decrease in hippocampal transient receptor potential M2 (TRPM2) channel and muscarinic acetylcholine receptor 1 (CHRM1) is associated with memory loss in a surgical menopause rat model

doi: 10.5114/aoms.2019.83760

Figure Lengend Snippet: CHRM1, TRPM2 immunoreactivity and apoptotic histoscore (diffuseness × severity) and MDA levels

Article Snippet: In order to prevent surface staining, after treatment with Ultra V Block (TA-125-UB, the Lab Vision Corporation, USA) solutions, the tissues were incubated with primary antibodies for 60 min (CHRM1 was purchased from Boster (Cholinergic receptor, muscarinic 1, catalog number: PA2202, Boster, 3942 B Valley Ave, Pleasanton, CA, 94566) and TRPM2 was purchased from Abcam (Rabbit Anti-TRPM2 antibody, ab101738, Abcam, Cambridge, UK)).

Techniques: TUNEL Assay

Journal: Archives of Medical Science : AMS

Article Title: The decrease in hippocampal transient receptor potential M2 (TRPM2) channel and muscarinic acetylcholine receptor 1 (CHRM1) is associated with memory loss in a surgical menopause rat model

doi: 10.5114/aoms.2019.83760

Figure Lengend Snippet: A – Increased chrm1 immunoreactivity staining in hippocampal area is shown in G1 relative to G2 (black arrow). B – Increased trpm2 immunoreactivity staining in hippocampal area is shown in G1 relative to G2 (black arrow). C – TUNEL staining in hippocampal area is lower in G1 compared with G2 (black arrow). D – CHRM1 immunoreactivity staining in hippocampal area is lower in G2 compared with G1 (black arrow). E – TRPM2 immunoreactivity staining in hippocampal area is lower in G2 compared with G1 (black arrow). F – TUNEL staining in hippocampal area is higher in G2 compared with G1 (black arrow)

Article Snippet: In order to prevent surface staining, after treatment with Ultra V Block (TA-125-UB, the Lab Vision Corporation, USA) solutions, the tissues were incubated with primary antibodies for 60 min (CHRM1 was purchased from Boster (Cholinergic receptor, muscarinic 1, catalog number: PA2202, Boster, 3942 B Valley Ave, Pleasanton, CA, 94566) and TRPM2 was purchased from Abcam (Rabbit Anti-TRPM2 antibody, ab101738, Abcam, Cambridge, UK)).

Techniques: Staining, TUNEL Assay

Journal: Molecules

Article Title: Detection of Biomarker Clusterin in SERS Immunoassays on Al Foil After Substrate Selection and Assay Optimization with Fluorescently Labeled Antibodies

doi: 10.3390/molecules30193974

Figure Lengend Snippet: ( A ) SERS spectra and ( B ) calibration plot in SERS immunoassay of clusterin at various concentrations of target clusterin on Al foil matte substrate, obtained with a 633 nm excitation laser. ( C ) Three- and ( D ) four-parameter logistic nonlinear regression analysis calibration plots for detection of clusterin in SERS sandwich immunoassay on Al foil matte substrate.

Article Snippet: Native human IgG protein (Abcam, AB205806 , Cambridge, Cambridgeshire, UK), anti-human IgG antibody produced from goat (H + L) coupled to ATTO647N (Hypermol), anti-human IgG antibody produced from goat (CSB_PA793692), and human clusterin protein (MW: 50.9 kDa) were obtained from ProSci Incorporated (Poway, CA, USA), while clusterin recombinant monoclonal antibody derived from human clusterin (CSB-RA449834A0HU) was purchased from Cusabio Technology LLC (Houston, TX, USA).

Techniques:

Journal: Cancer Science

Article Title: Immunophenotypic features of metastatic lymph node tumors to predict recurrence in N2 lung squamous cell carcinoma

doi: 10.1111/cas.12434

Figure Lengend Snippet: Univariate analysis of immunochemical staining of (a) cancer cells and (b) cancer-associated fibroblasts in primary tumors in patients with resected pathological N2 squamous cell carcinoma of the lung ( n = 64)

Article Snippet: Caveolin (clone D46G3; Cell Signaling, Danvers, MA, USA),( , ) clusterin (clone 1A11; Acris Antibodies, Herford, Germany),( , ) E-cadherin (clone 36; BD Biosciences, San Jose, CA, USA),( , ) and ZEB2 (Novus Biologicals, Littleton, CO, USA)( , ) were used as EMT-related markers.

Techniques: Staining, Variant Assay

Journal: Cancer Science

Article Title: Immunophenotypic features of metastatic lymph node tumors to predict recurrence in N2 lung squamous cell carcinoma

doi: 10.1111/cas.12434

Figure Lengend Snippet: (a) Univariate analysis of immunochemical staining of cancer cells in metastatic tumors in patients with resected pathological N2 squamous cell carcinoma of the lung ( n = 64); (b) Univariate analysis of immunochemical staining of cancer-associated fibroblasts in metastatic lymph node tumors in patients with resected pathological N2 squamous cell carcinoma of the lung

Article Snippet: Caveolin (clone D46G3; Cell Signaling, Danvers, MA, USA),( , ) clusterin (clone 1A11; Acris Antibodies, Herford, Germany),( , ) E-cadherin (clone 36; BD Biosciences, San Jose, CA, USA),( , ) and ZEB2 (Novus Biologicals, Littleton, CO, USA)( , ) were used as EMT-related markers.

Techniques: Staining, Variant Assay

Journal: Cancer Science

Article Title: Immunophenotypic features of metastatic lymph node tumors to predict recurrence in N2 lung squamous cell carcinoma

doi: 10.1111/cas.12434

Figure Lengend Snippet: Immunohistochemical staining of tumor cells in a primary tumor and a metastatic lymph node tumor in a patient with squamous cell carcinoma of the lung. (a,b) Clusterin expression of cancer cells in the primary tumor (a) and the metastatic lymph node tumor (b). (c,d) ZEB2 expression of cancer cells in the primary tumor (c) and the metastatic lymph node tumor (d). (e,f) Podoplanin expression of cancer-associated fibroblasts in the primary tumor (e) and the metastatic lymph node tumor (f). Dotted lines show the margin of the cancer cell nest.

Article Snippet: Caveolin (clone D46G3; Cell Signaling, Danvers, MA, USA),( , ) clusterin (clone 1A11; Acris Antibodies, Herford, Germany),( , ) E-cadherin (clone 36; BD Biosciences, San Jose, CA, USA),( , ) and ZEB2 (Novus Biologicals, Littleton, CO, USA)( , ) were used as EMT-related markers.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Cancer Science

Article Title: Immunophenotypic features of metastatic lymph node tumors to predict recurrence in N2 lung squamous cell carcinoma

doi: 10.1111/cas.12434

Figure Lengend Snippet: Kaplan–Meier recurrence-free survival (RFS) curve for patients with resected pathological N2 squamous cell carcinoma of the lung according to immunohistochemical staining. (a) Kaplan–Meier RFS curve according to clusterin expression of cancer cells in metastatic lymph node tumors. (b) Kaplan–Meier RFS curve according to ZEB2 expression of cancer cells in metastatic lymph node tumors. (c) Kaplan–Meier RFS curve according to podoplanin expression of cancer-associated fibroblasts in metastatic lymph node tumors.

Article Snippet: Caveolin (clone D46G3; Cell Signaling, Danvers, MA, USA),( , ) clusterin (clone 1A11; Acris Antibodies, Herford, Germany),( , ) E-cadherin (clone 36; BD Biosciences, San Jose, CA, USA),( , ) and ZEB2 (Novus Biologicals, Littleton, CO, USA)( , ) were used as EMT-related markers.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Cancer Science

Article Title: Immunophenotypic features of metastatic lymph node tumors to predict recurrence in N2 lung squamous cell carcinoma

doi: 10.1111/cas.12434

Figure Lengend Snippet: Multivariate analysis of clinicopathological factors for recurrence-free survival in patients with resected pathological N2 squamous cell carcinoma of the lung ( n = 64)

Article Snippet: Caveolin (clone D46G3; Cell Signaling, Danvers, MA, USA),( , ) clusterin (clone 1A11; Acris Antibodies, Herford, Germany),( , ) E-cadherin (clone 36; BD Biosciences, San Jose, CA, USA),( , ) and ZEB2 (Novus Biologicals, Littleton, CO, USA)( , ) were used as EMT-related markers.

Techniques: Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Protein levels of clusterin and glutathione synthetase in platelets allow for early detection of colorectal cancer

doi: 10.1007/s00018-017-2631-9

Figure Lengend Snippet: IPA networks of direct relationships. IPA networks of 14 proteins differentially expressed a between healthy controls vs. CRC patients (early- and late-stage) with a score of 40 and b between healthy controls vs. early-stage CRCs with a score of 41. Red marked proteins are lower expressed in CRC platelet samples compared to healthy controls, whereas green highlighted proteins are higher expressed in cancer samples. Out of these networks, clusterin (gene symbol: CLU) and glutathione synthetase (gene symbol: GSS) were selected for subsequent validation

Article Snippet: The membrane was blocked at room temperature for 1 h with 2% Amersham ECL Prime Blocking Agent (GE Healthcare), dissolved in 1× TBS with 0.1% Tween-20 (pH 7.6, Cell Signaling, USA) and incubated with primary antibodies against clusterin (rabbit monoclonal antibody, clone EPR2911, TA307501, OriGene), cofilin-1 (mouse monoclonal antibody, clone E-8, sc-376476, Santa Cruz Biotechnology, Inc., USA) and glutathione synthetase (rabbit polyclonal antibody, clone H-300, sc-28966, Santa Cruz Biotechnology) in 2% blocking buffer at 4 °C overnight.

Techniques:

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Protein levels of clusterin and glutathione synthetase in platelets allow for early detection of colorectal cancer

doi: 10.1007/s00018-017-2631-9

Figure Lengend Snippet: Western blot validation of platelet target proteins clusterin, GSH-S, and cofilin-1. Specific antibody-targeted protein bands were detected by Cy3-labeled secondary antibody. Cy5 total protein signals within each lane were used for normalization (Cy3/Cy5 ratio). Based on an internal standard, adjusted relative protein-level calculation was performed. Western blot validation of selected target proteins a clusterin, c GSH-S, and e cofilin-1 confirmed significant different level characteristics obtained by 2D-DIGE between distinct groups (p < 0.05). Based on ROC curve calculation, cut-offs for clusterin, GSH-S, and cofilin-1 were selected grouping values of protein levels into healthy control or early-stage CRC patient. Red lines represent median values. Maximal sensitivity is indicated by dashed lines and best specificity by dotted lines. a sensitivity; b specificity

Article Snippet: The membrane was blocked at room temperature for 1 h with 2% Amersham ECL Prime Blocking Agent (GE Healthcare), dissolved in 1× TBS with 0.1% Tween-20 (pH 7.6, Cell Signaling, USA) and incubated with primary antibodies against clusterin (rabbit monoclonal antibody, clone EPR2911, TA307501, OriGene), cofilin-1 (mouse monoclonal antibody, clone E-8, sc-376476, Santa Cruz Biotechnology, Inc., USA) and glutathione synthetase (rabbit polyclonal antibody, clone H-300, sc-28966, Santa Cruz Biotechnology) in 2% blocking buffer at 4 °C overnight.

Techniques: Western Blot, Labeling

Journal: PLoS ONE

Article Title: Common Human Cancer Genes Discovered by Integrated Gene-Expression Analysis

doi: 10.1371/journal.pone.0001149

Figure Lengend Snippet: Positive cytoplasmic staining of CLU presents on both tumor and normal cells in lung, breast, colon, and ovary tissue (A to D). CLU expression decreased in lung cancer (A, upper level) in comparison with normal lung (A, lower level). Both normal breast (B, lower level) and ovary (D, lower level) show middle to strong staining in cytoplasm, and middle to weak staining in breast (B, upper level) and ovarian cancer (D, upper level). Much less positive clusterin staining present in both colon tumor (C, upper level) and normal tissues (C, lower level). BID shows strong cytoplasmic and nuclear staining in lung cancer (E, upper level) and weak nuclear staining in normal lung epithelium (E, lower level). Increased strong nuclear and cytoplasmic staining is seen in breast tumor (F, upper level) when compared with normal breast tissue (F, lower level). In colon cancer, BID shows strong cytoplasmic and nuclear staining in tumor cells (G, upper level), while less positive BID staining was found in normal colon tissue (G, lower level). Semiquantitative analysis of CLU and BID immunoreaction in tumors and normal tissues (H). Most of samples from different types of cancer have strong, high percentage of BID immunoreaction, and normal tissues only show middle to low level staining; while CLU tends to down-regulated in tumors (H). High = score 10–12, middle = score 6–9, and low = score 1–5. Left column in each panel is at low power (100X) and right column in each panel is at high power (400X).

Article Snippet: After blocking in 10% of normal goat serum in PBS, all primary antibodies were incubated overnight at 4° C , including SPP1 (Novocastra Laboratories, Newcastle, UK, clone 15G12, dilution 1∶100), BID (BD Tranduction Laboratories, San Jose, CA, clone 7, dilution 1∶200) and CLU (Upstate Biotech, Lake Placid, NY, clone 41D, dilution 1∶1000).

Techniques: Staining, Expressing