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nuclear dye dapi  (Santa Cruz Biotechnology)


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    Structured Review

    Santa Cruz Biotechnology nuclear dye dapi
    p85α chaperones nuclear translocation <t>of</t> <t>OPN-i.</t> a, Immunoassay of lysates of 293T cells transfected with plasmids encoding Flag-p85α and OPN-i wild-type or OPN-i Y166F (YF) mutant, assessed by immunoprecipitation with anti-Flag and immunoblot analysis with anti-Flag and anti-OPN. Input, immunoblot analysis of an aliquot of cell lysates without IP. b, Confocal microscopy of OPN and Bcl-6 expression by CD44 + CD4 + T cells from OPN-i-KI mice 3 d post-injection with KLH in CFA, followed by cross-linking with anti-ICOS Ab (as in ) in vitro . Cells were counterstained with DNA-intercalating dye <t>DAPI</t> to trace nuclear perimeters. About 25–30 cells stained with Bcl-6 were further analyzed for localization of OPN protein. Right, fluorescence intensity expressed as the mean ratio of nuclear (Nuc) to cytoplasmic (Cyt) fluorescence pixel intensity ( n = 25–30 cells). c, Immunoblot analysis of nuclear fractions of OPN-i-KI CD62L − CD4 + T cells treated with anti-CD3 and/or anti-ICOS Abs (as in ), probed with anti-OPN, anti-Lamin B1 and anti-tubulin (to validate the integrity of nuclear separation). d , Immunoblot analysis of nuclear and cytosolic fractions of 293T cells transfected with plasmids encoding OPN-i wild-type or OPN-i Y166F mutant and increasing concentrations of Flag-p85α, probed with Abs, as indicated. e, Confocal microscopy of OPN and Bcl-6 expression by CD62L − CD4 + T cells from p85α WT or KO mice. Cell treatment and analysis as in a . Original magnification ( b, e ), 600×. * P < 0.05 and *** P < 0.001, Mann-Whitney test (error bars, mean ± s.e.m) (b, e ). All results are representative of at least two independent experiments.
    Nuclear Dye Dapi, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1459 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dapi+nuclear+dye+solution/DAPI/pmc04405167-188-26-17
    Average 96 stars, based on 1459 article reviews
    nuclear dye dapi - by Bioz Stars, 2026-10
    96/100 stars

    Images

    1) Product Images from "A p85α–osteopontin axis couples the ICOS receptor to sustained Bcl-6 expression by follicular helper and regulatory T cells"

    Article Title: A p85α–osteopontin axis couples the ICOS receptor to sustained Bcl-6 expression by follicular helper and regulatory T cells

    Journal: Nature immunology

    doi: 10.1038/ni.3050

    p85α chaperones nuclear translocation of OPN-i. a, Immunoassay of lysates of 293T cells transfected with plasmids encoding Flag-p85α and OPN-i wild-type or OPN-i Y166F (YF) mutant, assessed by immunoprecipitation with anti-Flag and immunoblot analysis with anti-Flag and anti-OPN. Input, immunoblot analysis of an aliquot of cell lysates without IP. b, Confocal microscopy of OPN and Bcl-6 expression by CD44 + CD4 + T cells from OPN-i-KI mice 3 d post-injection with KLH in CFA, followed by cross-linking with anti-ICOS Ab (as in ) in vitro . Cells were counterstained with DNA-intercalating dye DAPI to trace nuclear perimeters. About 25–30 cells stained with Bcl-6 were further analyzed for localization of OPN protein. Right, fluorescence intensity expressed as the mean ratio of nuclear (Nuc) to cytoplasmic (Cyt) fluorescence pixel intensity ( n = 25–30 cells). c, Immunoblot analysis of nuclear fractions of OPN-i-KI CD62L − CD4 + T cells treated with anti-CD3 and/or anti-ICOS Abs (as in ), probed with anti-OPN, anti-Lamin B1 and anti-tubulin (to validate the integrity of nuclear separation). d , Immunoblot analysis of nuclear and cytosolic fractions of 293T cells transfected with plasmids encoding OPN-i wild-type or OPN-i Y166F mutant and increasing concentrations of Flag-p85α, probed with Abs, as indicated. e, Confocal microscopy of OPN and Bcl-6 expression by CD62L − CD4 + T cells from p85α WT or KO mice. Cell treatment and analysis as in a . Original magnification ( b, e ), 600×. * P < 0.05 and *** P < 0.001, Mann-Whitney test (error bars, mean ± s.e.m) (b, e ). All results are representative of at least two independent experiments.
    Figure Legend Snippet: p85α chaperones nuclear translocation of OPN-i. a, Immunoassay of lysates of 293T cells transfected with plasmids encoding Flag-p85α and OPN-i wild-type or OPN-i Y166F (YF) mutant, assessed by immunoprecipitation with anti-Flag and immunoblot analysis with anti-Flag and anti-OPN. Input, immunoblot analysis of an aliquot of cell lysates without IP. b, Confocal microscopy of OPN and Bcl-6 expression by CD44 + CD4 + T cells from OPN-i-KI mice 3 d post-injection with KLH in CFA, followed by cross-linking with anti-ICOS Ab (as in ) in vitro . Cells were counterstained with DNA-intercalating dye DAPI to trace nuclear perimeters. About 25–30 cells stained with Bcl-6 were further analyzed for localization of OPN protein. Right, fluorescence intensity expressed as the mean ratio of nuclear (Nuc) to cytoplasmic (Cyt) fluorescence pixel intensity ( n = 25–30 cells). c, Immunoblot analysis of nuclear fractions of OPN-i-KI CD62L − CD4 + T cells treated with anti-CD3 and/or anti-ICOS Abs (as in ), probed with anti-OPN, anti-Lamin B1 and anti-tubulin (to validate the integrity of nuclear separation). d , Immunoblot analysis of nuclear and cytosolic fractions of 293T cells transfected with plasmids encoding OPN-i wild-type or OPN-i Y166F mutant and increasing concentrations of Flag-p85α, probed with Abs, as indicated. e, Confocal microscopy of OPN and Bcl-6 expression by CD62L − CD4 + T cells from p85α WT or KO mice. Cell treatment and analysis as in a . Original magnification ( b, e ), 600×. * P < 0.05 and *** P < 0.001, Mann-Whitney test (error bars, mean ± s.e.m) (b, e ). All results are representative of at least two independent experiments.

    Techniques Used: Translocation Assay, Transfection, Mutagenesis, Immunoprecipitation, Western Blot, Confocal Microscopy, Expressing, Injection, In Vitro, Staining, Fluorescence, MANN-WHITNEY

    Related Articles

    Staining:

    Article Title: Exposure of Human Corneal Epithelial Cells to Microplastic Particles Induces a Phase-Specific Cytokine Response.
    Article Snippet: 37 Microplastics have been identified in ophthalmic therapeutics and linked to dry eye 38 disease and meibomian gland dysfunction.. Inflammation is hypothesised to drive 39 ocular surface damage.. This in-vitro study investigates how varying concentrations of 40 microplastics affect corneal cell viability and cytokine expression.

    Article Title: CD200 + fibroblasts prime the immunoregulatory function of granulocytic myeloid-derived suppressor cells in inflammatory arthritis.
    Article Snippet: .. Secondary antibodies were stained for 1 h at room temperature followed by incubation with DAPI nuclear staining (1:1000, Cat# sc-3598, Santa Cruz). .. The following antibodies were used in human tissues: anti-CD33 (Polyclonal, 1:100, Cat# bs-2042R, Bioss), Nature Communications | (2026) 17:10746 12 anti-HLA-DR (LN3, 1:100, Cat# LS-B8114, LSBio), anti-CD20 (H1, 1:500, Cat# 555677, BD Biosciences), anti-CD56 (123A8, 1:1000, Cat# NBP244713, Novus), anti-CD11c (Polyclonal, 1:300, Cat# orb13554, Biorbyt), anti-CD3 (PS1, 1:50, Cat# ab699, Abcam), anti-CD14 (4B4F12, 1:100, Cat# ab182032, Abcam), anti-CD16 (2Q1240, 1:50, Cat# sc-70548, Santa Cruz), anti-CD123 (6H6, 1:100, Cat# 306002, BioLegend), anti-CD274/ PD-L1 (E1L3N, 1:100, Cat# 85164SF, Cell Signaling), anti-CD200 (Polyclonal, 1:100, Cat#AF2724, R&DSystems), anti-Podoplanin (Polyclonal, 1:100, Cat# AF3670, R&D Systems).

    Article Title: Macrophage-derived IL-6 reprograms lipid metabolism to promote colorectal cancer development through USP14-mediated FASN deubiquitination
    Article Snippet: For immunohistochemistry, the slices were incubated with antibodies against IL-6 (Cat#AF-206-NA, R&D Systems, Minneapolis, Minnesota, USA), p-STAT3 (Cell Signaling Technology, Danvers, Massachusetts, USA), USP14 (Proteintech, Wuhan, Hubei, China) and FASN (Cat#10624-2-AP, Proteintech) and then treated with an HRP-labeled secondary antibody (Proteintech). .. After the final round of staining, DAPI (Cat#sc-3598, Santa Cruz Biotechnology, Texas, USA) solution was added. .. Photomicrographs were obtained via a light microscope (Leica, Germany).

    Article Title: Glycine receptors in circulating white blood cells regulated by neuroinflammation
    Article Snippet: After incubation, the slides were washed three times with PBS for 5 minutes each, then incubated with appropriate secondary antibodies, i.e., Alexa Fluor 488 (1:200) and/or 594 (1:200) (Thermo-Fisher Scientific) for 1 hour at room temperature. .. A DAPI mounting agent (# AB104139 , Santa Cruz Biotechnology) was used for nuclear staining and mounting. ..

    Fluorescence:

    Article Title: Exposure of Human Corneal Epithelial Cells to Microplastic Particles Induces a Phase-Specific Cytokine Response.
    Article Snippet: 37 Microplastics have been identified in ophthalmic therapeutics and linked to dry eye 38 disease and meibomian gland dysfunction.. Inflammation is hypothesised to drive 39 ocular surface damage.. This in-vitro study investigates how varying concentrations of 40 microplastics affect corneal cell viability and cytokine expression.

    Microscopy:

    Article Title: Exposure of Human Corneal Epithelial Cells to Microplastic Particles Induces a Phase-Specific Cytokine Response.
    Article Snippet: 37 Microplastics have been identified in ophthalmic therapeutics and linked to dry eye 38 disease and meibomian gland dysfunction.. Inflammation is hypothesised to drive 39 ocular surface damage.. This in-vitro study investigates how varying concentrations of 40 microplastics affect corneal cell viability and cytokine expression.

    Incubation:

    Article Title: CD200 + fibroblasts prime the immunoregulatory function of granulocytic myeloid-derived suppressor cells in inflammatory arthritis.
    Article Snippet: .. Secondary antibodies were stained for 1 h at room temperature followed by incubation with DAPI nuclear staining (1:1000, Cat# sc-3598, Santa Cruz). .. The following antibodies were used in human tissues: anti-CD33 (Polyclonal, 1:100, Cat# bs-2042R, Bioss), Nature Communications | (2026) 17:10746 12 anti-HLA-DR (LN3, 1:100, Cat# LS-B8114, LSBio), anti-CD20 (H1, 1:500, Cat# 555677, BD Biosciences), anti-CD56 (123A8, 1:1000, Cat# NBP244713, Novus), anti-CD11c (Polyclonal, 1:300, Cat# orb13554, Biorbyt), anti-CD3 (PS1, 1:50, Cat# ab699, Abcam), anti-CD14 (4B4F12, 1:100, Cat# ab182032, Abcam), anti-CD16 (2Q1240, 1:50, Cat# sc-70548, Santa Cruz), anti-CD123 (6H6, 1:100, Cat# 306002, BioLegend), anti-CD274/ PD-L1 (E1L3N, 1:100, Cat# 85164SF, Cell Signaling), anti-CD200 (Polyclonal, 1:100, Cat#AF2724, R&DSystems), anti-Podoplanin (Polyclonal, 1:100, Cat# AF3670, R&D Systems).

    Article Title: Submedius thalamic inputs shape the presynaptic architecture of layer 5 orbitofrontal cortex in mice
    Article Snippet: For quantitative analysis of trans-synaptic tracing, 40-μm coronal sections of the whole brain were collected using cryostat (model #CM1860; Leica). .. Free-floating slices were incubated at room temperature with gentle agitation as follows: 2 h in blocking solution (5% heat-inactivated goat serum, 0.4% Triton X-100 in PBS); overnight in primary antibody diluted 1:1000 in blocking solution (mouse anti-GFP, GFP-1010, Aves Labs; or anti-RFP, 5f8, Chromotek); 2–3 h in secondary antibody diluted 1:500 (donkey anti-chicken-IgY Alexa488 or goat anti-rat IgG Cy3, Jackson ImmunoResearch); and 15 min in 2.5 μg/mL of DAPI (Santa Cruz, Cat #sc-3598) in PBS. .. Mice were anesthetized with 65 mg/kg ketamine (Daiichi-Sankyo) and 13 mg/kg xylazine (Sigma-Aldrich) via intraperitoneal injection and head-fixed in stereotaxic equipment (Narishige).

    Article Title: Donor-intrinsic regenerative potential outweighs histocompatibility in iPSC-derived skeletal muscle engraftment in nonhuman primates.
    Article Snippet: Cultured cells were washed with PBS and fixed with 4% paraformaldehyde (Millipore Sigma) for 15 min. .. Cells were then permeabilized with 0.3% Triton X-100 (Millipore Sigma) in PBS for 15 min at room temperature, blocked with 3% Bovine Serum Albumin (BSA) in PBS for 45 min, and incubated with primary antibodies overnight at 4 ◦ C. Following washing, cells were incubated with fluorophore-conjugated secondary antibodies containing DAPI (Santa Cruz, Dallas, TX, USA) for 1 h at room temperature. .. Mouse muscle cryosections were fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X-100 in PBS, blocked with 3% BSA, and incubated with primary antibodies overnight at 4 ◦ C. Sections were subsequently incubated with secondary antibodies and mounted using ProLong Gold Antifade Mountant with DAPI (Thermo Fisher Scientific).

    Gentle:

    Article Title: Submedius thalamic inputs shape the presynaptic architecture of layer 5 orbitofrontal cortex in mice
    Article Snippet: For quantitative analysis of trans-synaptic tracing, 40-μm coronal sections of the whole brain were collected using cryostat (model #CM1860; Leica). .. Free-floating slices were incubated at room temperature with gentle agitation as follows: 2 h in blocking solution (5% heat-inactivated goat serum, 0.4% Triton X-100 in PBS); overnight in primary antibody diluted 1:1000 in blocking solution (mouse anti-GFP, GFP-1010, Aves Labs; or anti-RFP, 5f8, Chromotek); 2–3 h in secondary antibody diluted 1:500 (donkey anti-chicken-IgY Alexa488 or goat anti-rat IgG Cy3, Jackson ImmunoResearch); and 15 min in 2.5 μg/mL of DAPI (Santa Cruz, Cat #sc-3598) in PBS. .. Mice were anesthetized with 65 mg/kg ketamine (Daiichi-Sankyo) and 13 mg/kg xylazine (Sigma-Aldrich) via intraperitoneal injection and head-fixed in stereotaxic equipment (Narishige).

    Blocking Assay:

    Article Title: Submedius thalamic inputs shape the presynaptic architecture of layer 5 orbitofrontal cortex in mice
    Article Snippet: For quantitative analysis of trans-synaptic tracing, 40-μm coronal sections of the whole brain were collected using cryostat (model #CM1860; Leica). .. Free-floating slices were incubated at room temperature with gentle agitation as follows: 2 h in blocking solution (5% heat-inactivated goat serum, 0.4% Triton X-100 in PBS); overnight in primary antibody diluted 1:1000 in blocking solution (mouse anti-GFP, GFP-1010, Aves Labs; or anti-RFP, 5f8, Chromotek); 2–3 h in secondary antibody diluted 1:500 (donkey anti-chicken-IgY Alexa488 or goat anti-rat IgG Cy3, Jackson ImmunoResearch); and 15 min in 2.5 μg/mL of DAPI (Santa Cruz, Cat #sc-3598) in PBS. .. Mice were anesthetized with 65 mg/kg ketamine (Daiichi-Sankyo) and 13 mg/kg xylazine (Sigma-Aldrich) via intraperitoneal injection and head-fixed in stereotaxic equipment (Narishige).

    FACS:

    Article Title: Combination therapy with targeted 4-1BB (CD137) agonists/anti-FAP binding domain and anti-CEA/anti-CD3 bispecific antibody
    Article Snippet: Cells were surface-stained with fluorescent dye-conjugated antibodies anti-human CD4 (clone RPA-T4, BioLegend, Cat.-No. 300532), CD8 (clone RPa-T8, BioLegend, Cat.-No. 3010441), CD62L (clone DREG-56, BioLegend, Cat.-No. 304834), CD127 (clone 019D5, BioLegend, Cat.-No. A019D5), CD134 (clone Ber-ACT35, BioLegend, Cat.-No. 350008), CD137 (clone 4B4-1, BioLegend, Cat.-No. 309814), GITR (clone 621, BioLegend, Cat.-No. 3311608) and CD25 (clone M-A251, BioLegend, Cat.-No. 356112) for 20 min at 4° C. in FACS buffer. .. Then, they were washed once with FACS buffer before being resuspended in 85 μL/well FACS buffer containing containing 0.2 μg/mL DAPI (Santa Cruz Biotec, Cat. No. Sc-3598) before they were acquired the same day using 5-laser LSR-Fortessa (BD Bioscience with DIVA software). ..

    Software:

    Article Title: Combination therapy with targeted 4-1BB (CD137) agonists/anti-FAP binding domain and anti-CEA/anti-CD3 bispecific antibody
    Article Snippet: Cells were surface-stained with fluorescent dye-conjugated antibodies anti-human CD4 (clone RPA-T4, BioLegend, Cat.-No. 300532), CD8 (clone RPa-T8, BioLegend, Cat.-No. 3010441), CD62L (clone DREG-56, BioLegend, Cat.-No. 304834), CD127 (clone 019D5, BioLegend, Cat.-No. A019D5), CD134 (clone Ber-ACT35, BioLegend, Cat.-No. 350008), CD137 (clone 4B4-1, BioLegend, Cat.-No. 309814), GITR (clone 621, BioLegend, Cat.-No. 3311608) and CD25 (clone M-A251, BioLegend, Cat.-No. 356112) for 20 min at 4° C. in FACS buffer. .. Then, they were washed once with FACS buffer before being resuspended in 85 μL/well FACS buffer containing containing 0.2 μg/mL DAPI (Santa Cruz Biotec, Cat. No. Sc-3598) before they were acquired the same day using 5-laser LSR-Fortessa (BD Bioscience with DIVA software). ..



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