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antifluorescence quenching blocking solution  (Beyotime)


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    Beyotime antifluorescence quenching blocking solution
    Antifluorescence Quenching Blocking Solution, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 2029 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/blocking+solution+containing+dapi/DAPI+Staining+Solution/pmc10053273-68-17-21
    Average 99 stars, based on 2029 article reviews
    antifluorescence quenching blocking solution - by Bioz Stars, 2026-09
    99/100 stars

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    Bicinchoninic Acid Protein Assay:

    Article Title: The host protein SSR4 mediates PRRSV-induced endoplasmic reticulum stress via interaction with Nsp2.
    Article Snippet: The following primary antibodies were used: anti-β-actin (Proteintech, 66009-1-Ig), anti-Calnexin (Proteintech, 10427-2-AP), anti-SSR1 (Proteintech, 10583-1-AP), anti-SSR2 (Proteintech, 10278-1-AP), anti-SSR3 (Proteintech, 30851-1-AP), anti-SSR4 (Proteintech, 11655-2-AP), anti-ATF4 (Proteintech, 10835-1-AP), anti-EDEM1 (Proteintech, 26226-1-AP), anti-GRP78 (Beyotime, AF0171), anti-ATF6 (Proteintech, 24169-1-AP), anti-Phospho-eIF2 alpha (Cell Signaling Technology, #3398), anti-eIF2 alpha (Cell Signaling Technol ogy, #5324), anti-Phospho-IRE1 (HUABIO, HA721980), anti-IRE1 (HUABIO, HA723225), anti-Puromycin (ABclonal, A21205), monoclonal anti-PRRSV N protein (generated in-house), and monoclonal anti-PRRSV Nsp2 (generated in-house). .. Key commercial reagents included tunicamycin (Beyotime, SC0393), DTT (Solarbio, 3483-12-3), 4-phenyl butyric acid (MCE, HY-A0281), MK-28 (AbMole, M56016), puromycin (MCE, HY-B1743), cycloheximide (MCE, HY-12320), BCA Protein Assay Kit (Thermo Fisher, 23225), Lipo fectamine 3000 (Thermo Fisher, L3000015), Lipofectamine RNAiMAX (Thermo Fisher, 13778075), ReverTra Ace qPCR RT Kit (TOYOBO, NLQ-101), KOD qPCR SYBR Mix (TOYOBO, QKD-201T), tunicamycin (MedChemExpress, HY-A0098), RIPA lysis buffer (Beyotime, P0013C), DAPI staining solution (Beyotime, C1006), and polyethylenimine (PEI; Yeasen). ..

    Article Title: Development of perilla essential oil-based nanoemulsions as anti-psoriatic and dermal delivery platforms for topical therapy
    Article Snippet: PEG-40 stearyl ether (S40) was obtained from Croda GmbH (Nett et al., Germany). .. DAPI staining solution, RIPA buffer, hematoxylin and eosin (H&E) staining kit, and bicinchoninic acid (BCA) protein assay kits were obtained from Beyotime Biotechnology (Shanghai, China). .. Antibodies against Ki67, TNF-α, IL-6, IL-1β, IL-17 and a one-step TUNEL assay kit were from Servicebio (Wuhan, China).

    Real-time Polymerase Chain Reaction:

    Article Title: The host protein SSR4 mediates PRRSV-induced endoplasmic reticulum stress via interaction with Nsp2.
    Article Snippet: The following primary antibodies were used: anti-β-actin (Proteintech, 66009-1-Ig), anti-Calnexin (Proteintech, 10427-2-AP), anti-SSR1 (Proteintech, 10583-1-AP), anti-SSR2 (Proteintech, 10278-1-AP), anti-SSR3 (Proteintech, 30851-1-AP), anti-SSR4 (Proteintech, 11655-2-AP), anti-ATF4 (Proteintech, 10835-1-AP), anti-EDEM1 (Proteintech, 26226-1-AP), anti-GRP78 (Beyotime, AF0171), anti-ATF6 (Proteintech, 24169-1-AP), anti-Phospho-eIF2 alpha (Cell Signaling Technology, #3398), anti-eIF2 alpha (Cell Signaling Technol ogy, #5324), anti-Phospho-IRE1 (HUABIO, HA721980), anti-IRE1 (HUABIO, HA723225), anti-Puromycin (ABclonal, A21205), monoclonal anti-PRRSV N protein (generated in-house), and monoclonal anti-PRRSV Nsp2 (generated in-house). .. Key commercial reagents included tunicamycin (Beyotime, SC0393), DTT (Solarbio, 3483-12-3), 4-phenyl butyric acid (MCE, HY-A0281), MK-28 (AbMole, M56016), puromycin (MCE, HY-B1743), cycloheximide (MCE, HY-12320), BCA Protein Assay Kit (Thermo Fisher, 23225), Lipo fectamine 3000 (Thermo Fisher, L3000015), Lipofectamine RNAiMAX (Thermo Fisher, 13778075), ReverTra Ace qPCR RT Kit (TOYOBO, NLQ-101), KOD qPCR SYBR Mix (TOYOBO, QKD-201T), tunicamycin (MedChemExpress, HY-A0098), RIPA lysis buffer (Beyotime, P0013C), DAPI staining solution (Beyotime, C1006), and polyethylenimine (PEI; Yeasen). ..

    Lysis:

    Article Title: The host protein SSR4 mediates PRRSV-induced endoplasmic reticulum stress via interaction with Nsp2.
    Article Snippet: The following primary antibodies were used: anti-β-actin (Proteintech, 66009-1-Ig), anti-Calnexin (Proteintech, 10427-2-AP), anti-SSR1 (Proteintech, 10583-1-AP), anti-SSR2 (Proteintech, 10278-1-AP), anti-SSR3 (Proteintech, 30851-1-AP), anti-SSR4 (Proteintech, 11655-2-AP), anti-ATF4 (Proteintech, 10835-1-AP), anti-EDEM1 (Proteintech, 26226-1-AP), anti-GRP78 (Beyotime, AF0171), anti-ATF6 (Proteintech, 24169-1-AP), anti-Phospho-eIF2 alpha (Cell Signaling Technology, #3398), anti-eIF2 alpha (Cell Signaling Technol ogy, #5324), anti-Phospho-IRE1 (HUABIO, HA721980), anti-IRE1 (HUABIO, HA723225), anti-Puromycin (ABclonal, A21205), monoclonal anti-PRRSV N protein (generated in-house), and monoclonal anti-PRRSV Nsp2 (generated in-house). .. Key commercial reagents included tunicamycin (Beyotime, SC0393), DTT (Solarbio, 3483-12-3), 4-phenyl butyric acid (MCE, HY-A0281), MK-28 (AbMole, M56016), puromycin (MCE, HY-B1743), cycloheximide (MCE, HY-12320), BCA Protein Assay Kit (Thermo Fisher, 23225), Lipo fectamine 3000 (Thermo Fisher, L3000015), Lipofectamine RNAiMAX (Thermo Fisher, 13778075), ReverTra Ace qPCR RT Kit (TOYOBO, NLQ-101), KOD qPCR SYBR Mix (TOYOBO, QKD-201T), tunicamycin (MedChemExpress, HY-A0098), RIPA lysis buffer (Beyotime, P0013C), DAPI staining solution (Beyotime, C1006), and polyethylenimine (PEI; Yeasen). ..

    Staining:

    Article Title: The host protein SSR4 mediates PRRSV-induced endoplasmic reticulum stress via interaction with Nsp2.
    Article Snippet: The following primary antibodies were used: anti-β-actin (Proteintech, 66009-1-Ig), anti-Calnexin (Proteintech, 10427-2-AP), anti-SSR1 (Proteintech, 10583-1-AP), anti-SSR2 (Proteintech, 10278-1-AP), anti-SSR3 (Proteintech, 30851-1-AP), anti-SSR4 (Proteintech, 11655-2-AP), anti-ATF4 (Proteintech, 10835-1-AP), anti-EDEM1 (Proteintech, 26226-1-AP), anti-GRP78 (Beyotime, AF0171), anti-ATF6 (Proteintech, 24169-1-AP), anti-Phospho-eIF2 alpha (Cell Signaling Technology, #3398), anti-eIF2 alpha (Cell Signaling Technol ogy, #5324), anti-Phospho-IRE1 (HUABIO, HA721980), anti-IRE1 (HUABIO, HA723225), anti-Puromycin (ABclonal, A21205), monoclonal anti-PRRSV N protein (generated in-house), and monoclonal anti-PRRSV Nsp2 (generated in-house). .. Key commercial reagents included tunicamycin (Beyotime, SC0393), DTT (Solarbio, 3483-12-3), 4-phenyl butyric acid (MCE, HY-A0281), MK-28 (AbMole, M56016), puromycin (MCE, HY-B1743), cycloheximide (MCE, HY-12320), BCA Protein Assay Kit (Thermo Fisher, 23225), Lipo fectamine 3000 (Thermo Fisher, L3000015), Lipofectamine RNAiMAX (Thermo Fisher, 13778075), ReverTra Ace qPCR RT Kit (TOYOBO, NLQ-101), KOD qPCR SYBR Mix (TOYOBO, QKD-201T), tunicamycin (MedChemExpress, HY-A0098), RIPA lysis buffer (Beyotime, P0013C), DAPI staining solution (Beyotime, C1006), and polyethylenimine (PEI; Yeasen). ..

    Article Title: A photodynamically activated nanoplatform relieves glucose-driven immunosuppression to potentiate STING immunotherapy in triple-negative breast cancer
    Article Snippet: Pre-screened foetal bovine serum (FBP-C520) was procured from Haixing Biosciences. .. DAPI Staining Solution, Cell Viability/Cytotoxicity assay kit, Cell Cycle and Apoptosis analysis kit, Reactive Oxygen Species (ROS) assay kit, and Mitochondrial Membrane Potential assay kit with JC-1 were all purchased from Beyotime, Inc. .. Cell Counting Kit-8 was obtained from TargetMol, USA.

    Article Title: Gastrodin alleviates neuronal damage in epileptic cell models by targeting P2RY12 to inhibit microglial hyperactivation
    Article Snippet: .. Then, DAPI staining solution (Beyotime, China, C1005, 1 μg/mL) was added, and the cells were incubated for 5 minutes to stain the nuclei. .. After washing with PBS, the images were acquired using a confocal microscope (Zeiss LSM880).

    Article Title: Targeted delivery of lupeol via hyaluronic acid-modified ZIF-8 for anti-hepatic fibrosis therapy
    Article Snippet: Quantitative analysis was performed using flow cytometry (Beckman Coulter, USA). .. A DAPI Staining Solution (Beyotime Biotechnology, Shanghai, China) was used to perform the cell apoptosis shape assay on LX-2 cells according to the manufacturer's instructions. .. Cells were seeded into six-well plates at a density of 1.0 × 10 6 cells per well and cultured for 24 h. The next day, cells were treated with different media containing Lupeol, Lupeol@ZIF-8 and HA/Lupeol@ZIF-8 (60 μM lupeol), with blank and model groups included as controls, and incubated for an additional 24 h. For qualitative analysis, DAPI solution was added 24 h after treatment, and fluorescence signals were recorded using a fluorescence microscope (Olympus, Tokyo, Japan).

    Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis
    Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China). .. The reagents used in the experiment included: H-DMEM(11965118, Gibco, USA.), α-DMEM medium(12571063, Gibco, USA.), TritonX-100(ST1723, Beyotime, China), 4 % paraformaldehyde (BL539A, Biosharp, China),FBS(A5256701, Gibco, USA.),ECM medium (Science Cell, USA.),and DAPI staining solution (C1006, Beyotime, China),BCIP/NBT(C3206, Beyotime, China), reactive oxygen species kit (S0033S, Beyotime, China), BSA (B2064, ≥98 %, Sigma-Aldrich, USA.),CD31 antibody (ab28364, Abcam, USA.), secondary anti-IGg (ab175773, Alexa Fluor® 680, Abcam, USA.), Phalloidin-iFluor 488(ab176753, Abcam, USA.), CCR7(AF5293, Bioss, China), CD206 (bsm-60761R, Bioss, China), iNOS (bs-22924R, Bioss, China), RIPA (P0013, Beyotime, China), p-ERK1/2 (AF3687, Affinity, USA.) and ERK1/2 (#AF0155, Affinity, USA), luminol detection reagent (sc-2048, Santa Cruz, USA.), GAPDH (Cat#KC-5G5, Kangchen Biotechnology, China), Trizol(15596026CN, Invitrogen, USA.), DEPC(R0601, Thermo Scientific, USA.), TBST(R017R.0000, Thermo Scientific, USA.), HIF-1a(GTX127309, GeneTex, USA.), β-Tubulin(10094-1-AP, Proteintech, UK) Adezmapimod (SB 203580, MCE, USA.) medium and Paclitaxel (99.88 %, HY-B0015R, MCE), ELISA Arg-1(E-EL-M3092, ELabSci@, China), TNF-α (E-EL-M3063, ELabSci@, China), OPN(22952-1-AP, Proteintech, UK.), F4/80 (GB11027-100, Servicebio, China) Alkaline phosphatase activity kit (P0321S, Beyotime, China), Matrigel (CLS356234, Corning, USA), Microfill MV120 (Flow tech, USA), EDTA(17892, Thermo Scientific, USA.) xylene(X112051, AR,99 %, Aladdin, China), ethanol (107-21-1, AR,99 %, Aladdin, China). .. All primers were designed and synthesized by Genema (Shanghai) Co., Ltd. 5.2.Fabrication of high-throughput array cell culture chip (HCGC) with Gaussian curvature gradient distribution. (1) Constructing models with different Gaussian curvatures: the multipipe of Grasshopper in Rhinocero software is used to regulate Gaussian curvature of the truss in the diamond framework in .

    Article Title: Development of perilla essential oil-based nanoemulsions as anti-psoriatic and dermal delivery platforms for topical therapy
    Article Snippet: PEG-40 stearyl ether (S40) was obtained from Croda GmbH (Nett et al., Germany). .. DAPI staining solution, RIPA buffer, hematoxylin and eosin (H&E) staining kit, and bicinchoninic acid (BCA) protein assay kits were obtained from Beyotime Biotechnology (Shanghai, China). .. Antibodies against Ki67, TNF-α, IL-6, IL-1β, IL-17 and a one-step TUNEL assay kit were from Servicebio (Wuhan, China).

    Article Title: Activation of the TGR5/cAMP/PKA/CREB axis in cholangiocytes mediates epithelial-mesenchymal transition and fibrosis in hepatolithiasis.
    Article Snippet: On the following day, cells were washed three times with PBS (5 minutes each), then 313 incubated at 37°C for 30 minutes with fluorescent secondary antibodies (e.g., Alexa Fluor 488/594 314 conjugates; Invitrogen, 1:500). .. After additional PBS washes, DAPI staining solution (Beyotime, 315 Cat# C1002, 1 μg/mL) was added and incubated in the dark for 3 minutes. ..

    Article Title: Gastrodin alleviates neuronal damage in epileptic cell models by targeting P2RY12 to inhibit microglial hyperactivation.
    Article Snippet: .. Then, DAPI staining solution (Beyotime, China, C1005, 1 μg/mL) was added, and the cells were incubated for 5 minutes to stain the nuclei. .. After washing with PBS, the images were acquired using a confocal microscope (Zeiss LSM880).

    ROS Assay:

    Article Title: A photodynamically activated nanoplatform relieves glucose-driven immunosuppression to potentiate STING immunotherapy in triple-negative breast cancer
    Article Snippet: Pre-screened foetal bovine serum (FBP-C520) was procured from Haixing Biosciences. .. DAPI Staining Solution, Cell Viability/Cytotoxicity assay kit, Cell Cycle and Apoptosis analysis kit, Reactive Oxygen Species (ROS) assay kit, and Mitochondrial Membrane Potential assay kit with JC-1 were all purchased from Beyotime, Inc. .. Cell Counting Kit-8 was obtained from TargetMol, USA.

    Membrane:

    Article Title: A photodynamically activated nanoplatform relieves glucose-driven immunosuppression to potentiate STING immunotherapy in triple-negative breast cancer
    Article Snippet: Pre-screened foetal bovine serum (FBP-C520) was procured from Haixing Biosciences. .. DAPI Staining Solution, Cell Viability/Cytotoxicity assay kit, Cell Cycle and Apoptosis analysis kit, Reactive Oxygen Species (ROS) assay kit, and Mitochondrial Membrane Potential assay kit with JC-1 were all purchased from Beyotime, Inc. .. Cell Counting Kit-8 was obtained from TargetMol, USA.

    Incubation:

    Article Title: Gastrodin alleviates neuronal damage in epileptic cell models by targeting P2RY12 to inhibit microglial hyperactivation
    Article Snippet: .. Then, DAPI staining solution (Beyotime, China, C1005, 1 μg/mL) was added, and the cells were incubated for 5 minutes to stain the nuclei. .. After washing with PBS, the images were acquired using a confocal microscope (Zeiss LSM880).

    Article Title: Activation of the TGR5/cAMP/PKA/CREB axis in cholangiocytes mediates epithelial-mesenchymal transition and fibrosis in hepatolithiasis.
    Article Snippet: On the following day, cells were washed three times with PBS (5 minutes each), then 313 incubated at 37°C for 30 minutes with fluorescent secondary antibodies (e.g., Alexa Fluor 488/594 314 conjugates; Invitrogen, 1:500). .. After additional PBS washes, DAPI staining solution (Beyotime, 315 Cat# C1002, 1 μg/mL) was added and incubated in the dark for 3 minutes. ..

    Article Title: Gastrodin alleviates neuronal damage in epileptic cell models by targeting P2RY12 to inhibit microglial hyperactivation.
    Article Snippet: .. Then, DAPI staining solution (Beyotime, China, C1005, 1 μg/mL) was added, and the cells were incubated for 5 minutes to stain the nuclei. .. After washing with PBS, the images were acquired using a confocal microscope (Zeiss LSM880).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis
    Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China). .. The reagents used in the experiment included: H-DMEM(11965118, Gibco, USA.), α-DMEM medium(12571063, Gibco, USA.), TritonX-100(ST1723, Beyotime, China), 4 % paraformaldehyde (BL539A, Biosharp, China),FBS(A5256701, Gibco, USA.),ECM medium (Science Cell, USA.),and DAPI staining solution (C1006, Beyotime, China),BCIP/NBT(C3206, Beyotime, China), reactive oxygen species kit (S0033S, Beyotime, China), BSA (B2064, ≥98 %, Sigma-Aldrich, USA.),CD31 antibody (ab28364, Abcam, USA.), secondary anti-IGg (ab175773, Alexa Fluor® 680, Abcam, USA.), Phalloidin-iFluor 488(ab176753, Abcam, USA.), CCR7(AF5293, Bioss, China), CD206 (bsm-60761R, Bioss, China), iNOS (bs-22924R, Bioss, China), RIPA (P0013, Beyotime, China), p-ERK1/2 (AF3687, Affinity, USA.) and ERK1/2 (#AF0155, Affinity, USA), luminol detection reagent (sc-2048, Santa Cruz, USA.), GAPDH (Cat#KC-5G5, Kangchen Biotechnology, China), Trizol(15596026CN, Invitrogen, USA.), DEPC(R0601, Thermo Scientific, USA.), TBST(R017R.0000, Thermo Scientific, USA.), HIF-1a(GTX127309, GeneTex, USA.), β-Tubulin(10094-1-AP, Proteintech, UK) Adezmapimod (SB 203580, MCE, USA.) medium and Paclitaxel (99.88 %, HY-B0015R, MCE), ELISA Arg-1(E-EL-M3092, ELabSci@, China), TNF-α (E-EL-M3063, ELabSci@, China), OPN(22952-1-AP, Proteintech, UK.), F4/80 (GB11027-100, Servicebio, China) Alkaline phosphatase activity kit (P0321S, Beyotime, China), Matrigel (CLS356234, Corning, USA), Microfill MV120 (Flow tech, USA), EDTA(17892, Thermo Scientific, USA.) xylene(X112051, AR,99 %, Aladdin, China), ethanol (107-21-1, AR,99 %, Aladdin, China). .. All primers were designed and synthesized by Genema (Shanghai) Co., Ltd. 5.2.Fabrication of high-throughput array cell culture chip (HCGC) with Gaussian curvature gradient distribution. (1) Constructing models with different Gaussian curvatures: the multipipe of Grasshopper in Rhinocero software is used to regulate Gaussian curvature of the truss in the diamond framework in .

    Activity Assay:

    Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis
    Article Snippet: The cells types utilized in this study included: hBMSCs (No. 7500, ScienCell, USA.), Raw264.7 (CL-0190, Procell system, China), HUVECs (No. 8000, ScienCell, USA.), rBMSCs (MUBMX-01001, OriCell, USA.) and MS1(CL-0162, Procell system, China). .. The reagents used in the experiment included: H-DMEM(11965118, Gibco, USA.), α-DMEM medium(12571063, Gibco, USA.), TritonX-100(ST1723, Beyotime, China), 4 % paraformaldehyde (BL539A, Biosharp, China),FBS(A5256701, Gibco, USA.),ECM medium (Science Cell, USA.),and DAPI staining solution (C1006, Beyotime, China),BCIP/NBT(C3206, Beyotime, China), reactive oxygen species kit (S0033S, Beyotime, China), BSA (B2064, ≥98 %, Sigma-Aldrich, USA.),CD31 antibody (ab28364, Abcam, USA.), secondary anti-IGg (ab175773, Alexa Fluor® 680, Abcam, USA.), Phalloidin-iFluor 488(ab176753, Abcam, USA.), CCR7(AF5293, Bioss, China), CD206 (bsm-60761R, Bioss, China), iNOS (bs-22924R, Bioss, China), RIPA (P0013, Beyotime, China), p-ERK1/2 (AF3687, Affinity, USA.) and ERK1/2 (#AF0155, Affinity, USA), luminol detection reagent (sc-2048, Santa Cruz, USA.), GAPDH (Cat#KC-5G5, Kangchen Biotechnology, China), Trizol(15596026CN, Invitrogen, USA.), DEPC(R0601, Thermo Scientific, USA.), TBST(R017R.0000, Thermo Scientific, USA.), HIF-1a(GTX127309, GeneTex, USA.), β-Tubulin(10094-1-AP, Proteintech, UK) Adezmapimod (SB 203580, MCE, USA.) medium and Paclitaxel (99.88 %, HY-B0015R, MCE), ELISA Arg-1(E-EL-M3092, ELabSci@, China), TNF-α (E-EL-M3063, ELabSci@, China), OPN(22952-1-AP, Proteintech, UK.), F4/80 (GB11027-100, Servicebio, China) Alkaline phosphatase activity kit (P0321S, Beyotime, China), Matrigel (CLS356234, Corning, USA), Microfill MV120 (Flow tech, USA), EDTA(17892, Thermo Scientific, USA.) xylene(X112051, AR,99 %, Aladdin, China), ethanol (107-21-1, AR,99 %, Aladdin, China). .. All primers were designed and synthesized by Genema (Shanghai) Co., Ltd. 5.2.Fabrication of high-throughput array cell culture chip (HCGC) with Gaussian curvature gradient distribution. (1) Constructing models with different Gaussian curvatures: the multipipe of Grasshopper in Rhinocero software is used to regulate Gaussian curvature of the truss in the diamond framework in .



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    90
    Beyotime antifluorescence quenching blocking solution containing dapi
    (A) The primary screen was performed on MCF10A cells in a 384-well format. Genes belonging to a subset of the human genome were knocked down in six replicate plates for 72 h by RNAi along with the listed controls. In a duplicate experiment, cells were stimulated with EGTA. Control wells were distributed on the plates as shown. NT: non-targeting KD – cells were transfected with non-targeting siRNA. No siRNA: cells received RNAiMAX only. Positive control (pos ctrl) – cells were transfected with siRNA against PSENEN. Transfection pos ctrl: cells were transfected with siRNA against polo-like kinase 1 (PLK1) as a cytotoxic indicator of transfection efficiency. Outer wells (yellow) were filled with media to prevent evaporation in experimental wells. (B) To assess endogenous NOTCH1 localization, cells in plates were subjected to automated IF <t>and</t> <t>DAPI/phalloidin</t> labeling to demarcate nuclei and the cell cortex, respectively. The script steps of the image acquisition and analysis developed in-house are as follows: (1) The nuclei were segmented based on DAPI signal. (2) The cell surface was segmented using the phalloidin signal and overlaid on the NOTCH1 channel (3) to quantify levels of cell surface NOTCH1 (4), and the area between the cell cortex and the nuclei was used to determine levels of cytoplasmic NOTCH1 (5). (6) The phalloidin mask was also used to count cells and establish cell-to-cell boundaries. (7) Nuclear size was used as readout of cell viability as compact; pyknotic nuclei identify dead cells. Using such a pipeline, each gene KD was defined by its effects on the amount of NOTCH1 in the cell cortex, in the cytoplasm or in the nucleus.
    Antifluorescence Quenching Blocking Solution Containing Dapi, supplied by Beyotime, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    blocking solution containing dapi  (Thermo Fisher)
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    Thermo Fisher blocking solution containing dapi
    Two-dimensional immunofluorescence stain of isolated bovine muscle satellite cells (BSCs). ( A ) Proliferating bovine satellite stained for <t>DAPI,</t> actin cytoskeleton (Phalloidin), and Pax7, a nuclear marker of satellite cells. Stains show a highly pure satellite cell population, following isolation and pre-plating protocol. ( B ) Following one week of differentiation, cells were stained for DAPI, actin cytoskeleton (Phalloidin), <t>and</t> <t>Troponin</t> T (CT3), a marker of myogenesis. Scale bars are 200 µm.
    Blocking Solution Containing Dapi, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/blocking+solution+containing+dapi/DyLight+755+Maleimide/pmc06835221-116-13-14
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    Image Search Results


    (A) The primary screen was performed on MCF10A cells in a 384-well format. Genes belonging to a subset of the human genome were knocked down in six replicate plates for 72 h by RNAi along with the listed controls. In a duplicate experiment, cells were stimulated with EGTA. Control wells were distributed on the plates as shown. NT: non-targeting KD – cells were transfected with non-targeting siRNA. No siRNA: cells received RNAiMAX only. Positive control (pos ctrl) – cells were transfected with siRNA against PSENEN. Transfection pos ctrl: cells were transfected with siRNA against polo-like kinase 1 (PLK1) as a cytotoxic indicator of transfection efficiency. Outer wells (yellow) were filled with media to prevent evaporation in experimental wells. (B) To assess endogenous NOTCH1 localization, cells in plates were subjected to automated IF and DAPI/phalloidin labeling to demarcate nuclei and the cell cortex, respectively. The script steps of the image acquisition and analysis developed in-house are as follows: (1) The nuclei were segmented based on DAPI signal. (2) The cell surface was segmented using the phalloidin signal and overlaid on the NOTCH1 channel (3) to quantify levels of cell surface NOTCH1 (4), and the area between the cell cortex and the nuclei was used to determine levels of cytoplasmic NOTCH1 (5). (6) The phalloidin mask was also used to count cells and establish cell-to-cell boundaries. (7) Nuclear size was used as readout of cell viability as compact; pyknotic nuclei identify dead cells. Using such a pipeline, each gene KD was defined by its effects on the amount of NOTCH1 in the cell cortex, in the cytoplasm or in the nucleus.

    Journal: Life Science Alliance

    Article Title: Novel determinants of NOTCH1 trafficking and signaling in breast epithelial cells

    doi: 10.26508/lsa.202403122

    Figure Lengend Snippet: (A) The primary screen was performed on MCF10A cells in a 384-well format. Genes belonging to a subset of the human genome were knocked down in six replicate plates for 72 h by RNAi along with the listed controls. In a duplicate experiment, cells were stimulated with EGTA. Control wells were distributed on the plates as shown. NT: non-targeting KD – cells were transfected with non-targeting siRNA. No siRNA: cells received RNAiMAX only. Positive control (pos ctrl) – cells were transfected with siRNA against PSENEN. Transfection pos ctrl: cells were transfected with siRNA against polo-like kinase 1 (PLK1) as a cytotoxic indicator of transfection efficiency. Outer wells (yellow) were filled with media to prevent evaporation in experimental wells. (B) To assess endogenous NOTCH1 localization, cells in plates were subjected to automated IF and DAPI/phalloidin labeling to demarcate nuclei and the cell cortex, respectively. The script steps of the image acquisition and analysis developed in-house are as follows: (1) The nuclei were segmented based on DAPI signal. (2) The cell surface was segmented using the phalloidin signal and overlaid on the NOTCH1 channel (3) to quantify levels of cell surface NOTCH1 (4), and the area between the cell cortex and the nuclei was used to determine levels of cytoplasmic NOTCH1 (5). (6) The phalloidin mask was also used to count cells and establish cell-to-cell boundaries. (7) Nuclear size was used as readout of cell viability as compact; pyknotic nuclei identify dead cells. Using such a pipeline, each gene KD was defined by its effects on the amount of NOTCH1 in the cell cortex, in the cytoplasm or in the nucleus.

    Article Snippet: Next, the cells were incubated for 1 h with 1% BSA blocking solution containing DAPI at 1:4,500 (Cat #: D9542; Merck Life Sciences), phalloidin at 1:350 (Cat #: P5282; Merck Life Sciences), and Alexa Fluor 488 anti-rat secondary antibody at 1:400 (Cat #: A21208; Thermo Fisher Scientific).

    Techniques: Control, Transfection, Positive Control, Evaporation, Labeling

    Two-dimensional immunofluorescence stain of isolated bovine muscle satellite cells (BSCs). ( A ) Proliferating bovine satellite stained for DAPI, actin cytoskeleton (Phalloidin), and Pax7, a nuclear marker of satellite cells. Stains show a highly pure satellite cell population, following isolation and pre-plating protocol. ( B ) Following one week of differentiation, cells were stained for DAPI, actin cytoskeleton (Phalloidin), and Troponin T (CT3), a marker of myogenesis. Scale bars are 200 µm.

    Journal: Foods

    Article Title: Extracellular Heme Proteins Influence Bovine Myosatellite Cell Proliferation and the Color of Cell-Based Meat

    doi: 10.3390/foods8100521

    Figure Lengend Snippet: Two-dimensional immunofluorescence stain of isolated bovine muscle satellite cells (BSCs). ( A ) Proliferating bovine satellite stained for DAPI, actin cytoskeleton (Phalloidin), and Pax7, a nuclear marker of satellite cells. Stains show a highly pure satellite cell population, following isolation and pre-plating protocol. ( B ) Following one week of differentiation, cells were stained for DAPI, actin cytoskeleton (Phalloidin), and Troponin T (CT3), a marker of myogenesis. Scale bars are 200 µm.

    Article Snippet: For BAMs, secondary antibodies for Troponin T were diluted in blocking solution containing DAPI (Thermo Fisher, #62248, 1:1000) and added to constructs for 1 h at room temperature.

    Techniques: Immunofluorescence, Staining, Isolation, Marker

    Confocal immunofluorescent imaging of BAMs. BAMs generated from BSC, BSC + Hb, or BSC + Mb (3 mg/mL for both heme proteins) were stained after eight days of differentiation for DAPI, actin cytoskeleton (Phalloidin), and Troponin T (CT3), a marker of myogenesis. Images show multinucleated myotube formation in BSC and BSC + Mb constructs, though not in BSC + Hb constructs. Scale bars are 200 µm.

    Journal: Foods

    Article Title: Extracellular Heme Proteins Influence Bovine Myosatellite Cell Proliferation and the Color of Cell-Based Meat

    doi: 10.3390/foods8100521

    Figure Lengend Snippet: Confocal immunofluorescent imaging of BAMs. BAMs generated from BSC, BSC + Hb, or BSC + Mb (3 mg/mL for both heme proteins) were stained after eight days of differentiation for DAPI, actin cytoskeleton (Phalloidin), and Troponin T (CT3), a marker of myogenesis. Images show multinucleated myotube formation in BSC and BSC + Mb constructs, though not in BSC + Hb constructs. Scale bars are 200 µm.

    Article Snippet: For BAMs, secondary antibodies for Troponin T were diluted in blocking solution containing DAPI (Thermo Fisher, #62248, 1:1000) and added to constructs for 1 h at room temperature.

    Techniques: Imaging, Generated, Staining, Marker, Construct