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pbs  (Thermo Fisher)


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    Structured Review

    Thermo Fisher pbs
    Pbs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/microarray+blocking+solution/Tween+20/pmc03484980-234-46-38
    Average 99 stars, based on 1 article reviews
    pbs - by Bioz Stars, 2026-10
    99/100 stars

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    Related Articles

    Membrane:

    Article Title: PIK3CA mutant cervical cancer is selectively suppressed by PI3Kα inhibition (Alpelisib/BYL-719 and Inavolisib/GDC-0077) and cooperates with HPV directed T cell therapy
    Article Snippet: According to the manufacturer's protocols, whole-cell protein lysates were prepared from cell lines using RIPA lysis buffer supplemented with Complete Protease Inhibitor Cocktail (Santa Cruz, CA, United States). .. In total, 20 - 40 micrograms of whole cell protein were separated in a 4–12% NuPAGE Bis-Tris-gel, transferred to PVDF membrane (Thermo Fisher Scientific, Carlsbad, CA, United States), and immunoblotted with primary antibodies in 0.05% Tween 20-Tris-buffered saline (TBST) containing 5% BSA at 4°C with shaking overnight. .. The primary antibodies used in this study were: rabbit monoclonal anti-PD-L1, (Cell Signaling Technology Cat# 13684, RRID: AB_2687655), YAP1, (Cell Signaling Technology Cat# 14074, RRID: AB_2650491), EGFR (Cell Signaling Technology Cat# 4267, RRID: AB_2800085), CTGF (Cell Signaling Technology Cat# 86641, RRID: AB_2800085), Integrin α6 (Cell Signaling Technology Cat# 3750, RRID: AB_2249263), IRF1 (Cell Signaling Technology Cat# 8478, RRID: AB_10949108) at a dilution from1:250 to 1:500; mouse monoclonal anti-HPV16 E7 at 1:100 (Santa Cruz Cat# sc-51951, RRID: AB_629662), and goat monoclonal anti-p16 at 1:500 (R&D system Cat# AF5779, RRID: AB_1964666). β-actin, used as an internal control, was detected by rabbit anti-β-actin antibody (Cell Signaling Technology Cat# 4967, RRID: AB_330288).

    Saline:

    Article Title: PIK3CA mutant cervical cancer is selectively suppressed by PI3Kα inhibition (Alpelisib/BYL-719 and Inavolisib/GDC-0077) and cooperates with HPV directed T cell therapy
    Article Snippet: According to the manufacturer's protocols, whole-cell protein lysates were prepared from cell lines using RIPA lysis buffer supplemented with Complete Protease Inhibitor Cocktail (Santa Cruz, CA, United States). .. In total, 20 - 40 micrograms of whole cell protein were separated in a 4–12% NuPAGE Bis-Tris-gel, transferred to PVDF membrane (Thermo Fisher Scientific, Carlsbad, CA, United States), and immunoblotted with primary antibodies in 0.05% Tween 20-Tris-buffered saline (TBST) containing 5% BSA at 4°C with shaking overnight. .. The primary antibodies used in this study were: rabbit monoclonal anti-PD-L1, (Cell Signaling Technology Cat# 13684, RRID: AB_2687655), YAP1, (Cell Signaling Technology Cat# 14074, RRID: AB_2650491), EGFR (Cell Signaling Technology Cat# 4267, RRID: AB_2800085), CTGF (Cell Signaling Technology Cat# 86641, RRID: AB_2800085), Integrin α6 (Cell Signaling Technology Cat# 3750, RRID: AB_2249263), IRF1 (Cell Signaling Technology Cat# 8478, RRID: AB_10949108) at a dilution from1:250 to 1:500; mouse monoclonal anti-HPV16 E7 at 1:100 (Santa Cruz Cat# sc-51951, RRID: AB_629662), and goat monoclonal anti-p16 at 1:500 (R&D system Cat# AF5779, RRID: AB_1964666). β-actin, used as an internal control, was detected by rabbit anti-β-actin antibody (Cell Signaling Technology Cat# 4967, RRID: AB_330288).

    Article Title: Adaptable sliding hydrogels enable pericellular pocket formation while enhancing MSC chondrogenesis and survival in 3D
    Article Snippet: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to separate lysates with 4-12% Bis-Tris gels (Thermo Scientific) before transfer to PVDF membranes using an iBlot3 (Invitrogen). .. Blocking with 5% BSA in Tris-buffered saline with Tween 20 (TBST) for 1 h at room temperature was followed by overnight incubation with primary antibodies diluted in SuperBlock blocking buffers (Thermo Fisher Scientific) at 4 °C. ..

    Lysis:

    Article Title: Dual role of ZIC2 during neural induction: from priming transcription factor to enhancer activator
    Article Snippet: .. Cells were resuspended in Cell Lysis Buffer [25 mM KCl (Sigma–Aldrich, 27810.295), 5 mM TRIS (Sigma–Aldrich, 0497–5KG) pH 8.3, 1.25 mM MgCl 2 (VWR BDH7899-1), 0.225% IGEPAL (Sigma–Aldrich, I8896-50 ML), and 0.225% Tween20 (VWR)] supplemented with 0.4 μg/μl Proteinase K (Thermo Fisher Scientific, EO0492), vortexed, and incubated sequentially at 65°C for 6 min and 95°C for 2 min. Lysates were cooled down, and 1–2 μl were used directly for polymerase chain reaction (PCR). .. For applications requiring high-purity gDNA, including Sanger sequencing, gDNA was isolated using the NZY Tissue gDNA Isolation Kit (NZYTech, MB13503) according to the manufacturer’s instructions.

    Incubation:

    Article Title: Dual role of ZIC2 during neural induction: from priming transcription factor to enhancer activator
    Article Snippet: .. Cells were resuspended in Cell Lysis Buffer [25 mM KCl (Sigma–Aldrich, 27810.295), 5 mM TRIS (Sigma–Aldrich, 0497–5KG) pH 8.3, 1.25 mM MgCl 2 (VWR BDH7899-1), 0.225% IGEPAL (Sigma–Aldrich, I8896-50 ML), and 0.225% Tween20 (VWR)] supplemented with 0.4 μg/μl Proteinase K (Thermo Fisher Scientific, EO0492), vortexed, and incubated sequentially at 65°C for 6 min and 95°C for 2 min. Lysates were cooled down, and 1–2 μl were used directly for polymerase chain reaction (PCR). .. For applications requiring high-purity gDNA, including Sanger sequencing, gDNA was isolated using the NZY Tissue gDNA Isolation Kit (NZYTech, MB13503) according to the manufacturer’s instructions.

    Article Title: Distinct hyaluronan content and hierarchical organization of collagen-elastic fiber assemblies in subcutaneous fascia
    Article Snippet: .. Samples were incubated with elastin monoclonal antibody BA-4 (MA1-27129, Thermo Fisher, Waltham, MA, USA; dilution 1: 100 in T-PBS containing 0.1% BSA) for 1 h at room temperature, washed three times for 5 min each with T-PBS (0.1% Tween-20 in PBS), and then reacted with Alexa Fluor 594–conjugated goat anti-mouse IgG (A11032; Thermo Fisher; dilution 1:1000 in T-PBS containing 0.1% BSA) for 1 h at room temperature [ ]. .. According to the manufacturer's protocol, 20 μM CHP (FLU60; 3Helix/Funakoshi) dissolved in PBS was heated at 80 °C for 5 min, rapidly cooled in an ice bath, and centrifuged; the supernatant was then used without further dilution [ , , ].

    Article Title: Silencing of SIVA-1 promotes cisplatin resistance in gastric cancer via the Bcl-2/BAX-mediated mitochondria-dependent apoptosis pathway
    Article Snippet: .. Subsequently, Goat Anti-Rabbit IgG H&L (HRP) (cat. no. ab6721; Abcam) was added and incubated for 1 h at room temperature in the dark, and washed three times with 0.1% TBS Tween-20 Buffer (cat. no. 28360; Thermo Fisher Scientific, Inc.). .. A gel imager (iBright CL750; Invitrogen; Thermo Fisher Scientific, Inc.) was used to scan the protein blotting membrane, and the gray scale values of the bands were analyzed and recorded using ImageJ software (bundled with 64-bit Java 8; National Institutes of Health).

    Article Title: Adaptable sliding hydrogels enable pericellular pocket formation while enhancing MSC chondrogenesis and survival in 3D
    Article Snippet: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to separate lysates with 4-12% Bis-Tris gels (Thermo Scientific) before transfer to PVDF membranes using an iBlot3 (Invitrogen). .. Blocking with 5% BSA in Tris-buffered saline with Tween 20 (TBST) for 1 h at room temperature was followed by overnight incubation with primary antibodies diluted in SuperBlock blocking buffers (Thermo Fisher Scientific) at 4 °C. ..

    Polymerase Chain Reaction:

    Article Title: Dual role of ZIC2 during neural induction: from priming transcription factor to enhancer activator
    Article Snippet: .. Cells were resuspended in Cell Lysis Buffer [25 mM KCl (Sigma–Aldrich, 27810.295), 5 mM TRIS (Sigma–Aldrich, 0497–5KG) pH 8.3, 1.25 mM MgCl 2 (VWR BDH7899-1), 0.225% IGEPAL (Sigma–Aldrich, I8896-50 ML), and 0.225% Tween20 (VWR)] supplemented with 0.4 μg/μl Proteinase K (Thermo Fisher Scientific, EO0492), vortexed, and incubated sequentially at 65°C for 6 min and 95°C for 2 min. Lysates were cooled down, and 1–2 μl were used directly for polymerase chain reaction (PCR). .. For applications requiring high-purity gDNA, including Sanger sequencing, gDNA was isolated using the NZY Tissue gDNA Isolation Kit (NZYTech, MB13503) according to the manufacturer’s instructions.

    Blocking Assay:

    Article Title: Adaptable sliding hydrogels enable pericellular pocket formation while enhancing MSC chondrogenesis and survival in 3D
    Article Snippet: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to separate lysates with 4-12% Bis-Tris gels (Thermo Scientific) before transfer to PVDF membranes using an iBlot3 (Invitrogen). .. Blocking with 5% BSA in Tris-buffered saline with Tween 20 (TBST) for 1 h at room temperature was followed by overnight incubation with primary antibodies diluted in SuperBlock blocking buffers (Thermo Fisher Scientific) at 4 °C. ..



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    Gsk3 negatively regulates mitotic effectors in Xenopus extracts. (A) Scheme of the protein <t>microarray</t> experiment to assess Gsk3-dependent polyubiquitination. Interphase egg extracts were supplemented with ubiquitin, pretreated with LiCl or NaCl (Control), and then applied on protein arrays containing ∼9,000 proteins. Polyubiquitination of arrayed proteins (PolyUb) was monitored using anti-polyubiquitin antibodies and immunofluorescence detection using an array scanner. Signal intensities were transformed and are displayed as heat-map. (B) Distribution plot of polyubiquitination changes of 3,873 in vitro polyubiquitinated proteins after Gsk3 inhibition by LiCl (Log2[Control/LiCl]). Arrays were treated with 30 mM LiCl or 30 mM NaCl (Control). LiCl reduced polyubiquitination of 864 proteins >1.8-fold. Some candidates, associated with cell cycle progression and selected for further validation are indicated in the plot. (C) Cluster of mitotic associated proteins that are significantly regulated by LiCl in B. The functional network was determined using String 9.1 and color-coded based on the fold change in the microarray (Dataset S1). (D) Western blots of indicated proteins from Xenopus eggs (naturally arrested in metaphase II) and released into interphase with the Ca2+ ionophore A23187 for 25 min, treated with LiCl, or mock treated (NaCl). β-cat, β-catenin; cycE, cyclin E; Interph., interphase; M. II, metaphase II; pH3, phosphorylated histone 3. (E) Western blots of stage VI oocytes in Prophase I and Progesterone-matured (Prog.) oocytes in metaphase II. Oocytes were injected (Inj.) with Morpholinos (MO) or mRNA as indicated. Co, control; dnWnt11, dominant negative Wnt11; pH3, phosphorylated histone 3; PPL, preprolactin (negative control).
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    Image Search Results


    Gsk3 negatively regulates mitotic effectors in Xenopus extracts. (A) Scheme of the protein microarray experiment to assess Gsk3-dependent polyubiquitination. Interphase egg extracts were supplemented with ubiquitin, pretreated with LiCl or NaCl (Control), and then applied on protein arrays containing ∼9,000 proteins. Polyubiquitination of arrayed proteins (PolyUb) was monitored using anti-polyubiquitin antibodies and immunofluorescence detection using an array scanner. Signal intensities were transformed and are displayed as heat-map. (B) Distribution plot of polyubiquitination changes of 3,873 in vitro polyubiquitinated proteins after Gsk3 inhibition by LiCl (Log2[Control/LiCl]). Arrays were treated with 30 mM LiCl or 30 mM NaCl (Control). LiCl reduced polyubiquitination of 864 proteins >1.8-fold. Some candidates, associated with cell cycle progression and selected for further validation are indicated in the plot. (C) Cluster of mitotic associated proteins that are significantly regulated by LiCl in B. The functional network was determined using String 9.1 and color-coded based on the fold change in the microarray (Dataset S1). (D) Western blots of indicated proteins from Xenopus eggs (naturally arrested in metaphase II) and released into interphase with the Ca2+ ionophore A23187 for 25 min, treated with LiCl, or mock treated (NaCl). β-cat, β-catenin; cycE, cyclin E; Interph., interphase; M. II, metaphase II; pH3, phosphorylated histone 3. (E) Western blots of stage VI oocytes in Prophase I and Progesterone-matured (Prog.) oocytes in metaphase II. Oocytes were injected (Inj.) with Morpholinos (MO) or mRNA as indicated. Co, control; dnWnt11, dominant negative Wnt11; pH3, phosphorylated histone 3; PPL, preprolactin (negative control).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Maternal Wnt/STOP signaling promotes cell division during early Xenopus embryogenesis

    doi: 10.1073/pnas.1423533112

    Figure Lengend Snippet: Gsk3 negatively regulates mitotic effectors in Xenopus extracts. (A) Scheme of the protein microarray experiment to assess Gsk3-dependent polyubiquitination. Interphase egg extracts were supplemented with ubiquitin, pretreated with LiCl or NaCl (Control), and then applied on protein arrays containing ∼9,000 proteins. Polyubiquitination of arrayed proteins (PolyUb) was monitored using anti-polyubiquitin antibodies and immunofluorescence detection using an array scanner. Signal intensities were transformed and are displayed as heat-map. (B) Distribution plot of polyubiquitination changes of 3,873 in vitro polyubiquitinated proteins after Gsk3 inhibition by LiCl (Log2[Control/LiCl]). Arrays were treated with 30 mM LiCl or 30 mM NaCl (Control). LiCl reduced polyubiquitination of 864 proteins >1.8-fold. Some candidates, associated with cell cycle progression and selected for further validation are indicated in the plot. (C) Cluster of mitotic associated proteins that are significantly regulated by LiCl in B. The functional network was determined using String 9.1 and color-coded based on the fold change in the microarray (Dataset S1). (D) Western blots of indicated proteins from Xenopus eggs (naturally arrested in metaphase II) and released into interphase with the Ca2+ ionophore A23187 for 25 min, treated with LiCl, or mock treated (NaCl). β-cat, β-catenin; cycE, cyclin E; Interph., interphase; M. II, metaphase II; pH3, phosphorylated histone 3. (E) Western blots of stage VI oocytes in Prophase I and Progesterone-matured (Prog.) oocytes in metaphase II. Oocytes were injected (Inj.) with Morpholinos (MO) or mRNA as indicated. Co, control; dnWnt11, dominant negative Wnt11; pH3, phosphorylated histone 3; PPL, preprolactin (negative control).

    Article Snippet: Briefly, the arrays were washed in TBST and blocked for 4 h at 4 °C using Microarray Blocking solution (Arrayit), followed by a wash in lysis buffer.

    Techniques: Microarray, Ubiquitin Proteomics, Control, Immunofluorescence, Transformation Assay, In Vitro, Inhibition, Biomarker Discovery, Functional Assay, Western Blot, Injection, Dominant Negative Mutation, Negative Control