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4 6 diamidino 2 phenylindole  (Dojindo Labs)


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    Structured Review

    Dojindo Labs 4 6 diamidino 2 phenylindole
    BBB maturation stages in the postnatal cerebral cortex based on glial configuration. (A) 3D images of AQP4, Iba1, lectin, and DAPI staining in the rat cerebral cortex at P1–30 are shown. Scale bar indicates 30 μm. AQP4 gradually accumulated in the blood vessels and localized to the blood vessels after P15. The number of Iba1+ microglia significantly increased after P10 and peaked at P15, changing their shape from ameboid to ramified. (B) Temporal changes in biotin permeability are corrected to a maximum value of 100%. Typical images of P4 and P15 rat brain cortical regions stained with streptavidin are shown. A solid line was drawn perpendicular to the blood vessel to quantify the fluorescence, which is indicated by the dashed line, and the red fluorescence value was measured, as shown in the upper right graph. The traces show the average of 70–76 transverse vessel lines for the respective postnatal ages. The maximum fluorescence value was set at 100, and the sum of the fluorescence values 20 μm to the left and to the right was calculated. The peak became sharper as the age increased. Astrocyte and microglial coverage rates were corrected to a P30 value of 100%. BBB formation and maturation in rats are classified into three phases based on biotin leakage and astrocyte and microglial contact with blood vessels . The phases are as follows: the “immature stage” when biotin permeability is still high; the “organic formation stage” when glial structures around blood vessels are formed; and the “completion stage” when the structures around blood vessels become stable and the morphology of microglia changes to a ramified type. Abbreviations: AQP4: aquaporin 4; BBB: blood–brain barrier; DAPI: <t>4′,6-diamidino-2-phenylindole;</t> Iba1: ionized calcium binding adapter protein 1; P: postnatal day.
    4 6 Diamidino 2 Phenylindole, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 95/100, based on 118 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dapi/Cellstain+DAPI/pmc12971911-28-16-20
    Average 95 stars, based on 118 article reviews
    4 6 diamidino 2 phenylindole - by Bioz Stars, 2026-09
    95/100 stars

    Images

    1) Product Images from "Search for marker proteins to assess blood–brain barrier development"

    Article Title: Search for marker proteins to assess blood–brain barrier development

    Journal: Frontiers in Neuroanatomy

    doi: 10.3389/fnana.2026.1717532

    BBB maturation stages in the postnatal cerebral cortex based on glial configuration. (A) 3D images of AQP4, Iba1, lectin, and DAPI staining in the rat cerebral cortex at P1–30 are shown. Scale bar indicates 30 μm. AQP4 gradually accumulated in the blood vessels and localized to the blood vessels after P15. The number of Iba1+ microglia significantly increased after P10 and peaked at P15, changing their shape from ameboid to ramified. (B) Temporal changes in biotin permeability are corrected to a maximum value of 100%. Typical images of P4 and P15 rat brain cortical regions stained with streptavidin are shown. A solid line was drawn perpendicular to the blood vessel to quantify the fluorescence, which is indicated by the dashed line, and the red fluorescence value was measured, as shown in the upper right graph. The traces show the average of 70–76 transverse vessel lines for the respective postnatal ages. The maximum fluorescence value was set at 100, and the sum of the fluorescence values 20 μm to the left and to the right was calculated. The peak became sharper as the age increased. Astrocyte and microglial coverage rates were corrected to a P30 value of 100%. BBB formation and maturation in rats are classified into three phases based on biotin leakage and astrocyte and microglial contact with blood vessels . The phases are as follows: the “immature stage” when biotin permeability is still high; the “organic formation stage” when glial structures around blood vessels are formed; and the “completion stage” when the structures around blood vessels become stable and the morphology of microglia changes to a ramified type. Abbreviations: AQP4: aquaporin 4; BBB: blood–brain barrier; DAPI: 4′,6-diamidino-2-phenylindole; Iba1: ionized calcium binding adapter protein 1; P: postnatal day.
    Figure Legend Snippet: BBB maturation stages in the postnatal cerebral cortex based on glial configuration. (A) 3D images of AQP4, Iba1, lectin, and DAPI staining in the rat cerebral cortex at P1–30 are shown. Scale bar indicates 30 μm. AQP4 gradually accumulated in the blood vessels and localized to the blood vessels after P15. The number of Iba1+ microglia significantly increased after P10 and peaked at P15, changing their shape from ameboid to ramified. (B) Temporal changes in biotin permeability are corrected to a maximum value of 100%. Typical images of P4 and P15 rat brain cortical regions stained with streptavidin are shown. A solid line was drawn perpendicular to the blood vessel to quantify the fluorescence, which is indicated by the dashed line, and the red fluorescence value was measured, as shown in the upper right graph. The traces show the average of 70–76 transverse vessel lines for the respective postnatal ages. The maximum fluorescence value was set at 100, and the sum of the fluorescence values 20 μm to the left and to the right was calculated. The peak became sharper as the age increased. Astrocyte and microglial coverage rates were corrected to a P30 value of 100%. BBB formation and maturation in rats are classified into three phases based on biotin leakage and astrocyte and microglial contact with blood vessels . The phases are as follows: the “immature stage” when biotin permeability is still high; the “organic formation stage” when glial structures around blood vessels are formed; and the “completion stage” when the structures around blood vessels become stable and the morphology of microglia changes to a ramified type. Abbreviations: AQP4: aquaporin 4; BBB: blood–brain barrier; DAPI: 4′,6-diamidino-2-phenylindole; Iba1: ionized calcium binding adapter protein 1; P: postnatal day.

    Techniques Used: Staining, Permeability, Fluorescence, Binding Assay

    Expression of blood vessel developmental markers in the postnatal cerebral cortex. (A) Images of CD31, CD34, CD146, agrin, and Tie2 co-stained with lectin and DAPI in the rat cerebral cortex at P1–30 are shown. Scale bar indicates 50 μm. CD31 and Tie2 signals gradually increased in blood vessels, whereas CD34, CD146, and agrin signals gradually decreased. (B) The graphs show temporal changes in the percentage of marker+ area relative to the total vessel area. The colocalized signals between the markers and blood vessels were calculated, and the areas of the colocalized signals were normalized to the total blood vessel area in each image. Approximately 9–15 Images were obtained from the cerebral cortical regions of three rat pups of each age with a Nikon A1R-A1 confocal microscope. Data are shown as averaged value ± SEM. Data were analyzed using an ANOVA followed by a Tukey’s multiple range test. * p < 0.05, ** p < 0.01. vs. P1 value, while # p < 0.05, ## p < 0.01 vs. P10 value. (C) Graph summarizing results for B and B. Changes in biotin permeability, CD34, CD146, and agrin were corrected to a P1 value of 100%, and the astrocyte coverage rate, microglial coverage rate, CD31, and Tie2 were corrected to a P30 value of 100%. The grey zone represents the organic formation term (P4P15). Abbreviations: CD31, cluster of differentiation 31 or platelet endothelial cell adhesion molecule 1; CD146, melanoma cell adhesion molecule; DAPI: 4′,6-diamidino-2-phenylindole; P, postnatal day; Tie2, TEK receptor tyrosine kinase; ANOVA, analysis of variance.
    Figure Legend Snippet: Expression of blood vessel developmental markers in the postnatal cerebral cortex. (A) Images of CD31, CD34, CD146, agrin, and Tie2 co-stained with lectin and DAPI in the rat cerebral cortex at P1–30 are shown. Scale bar indicates 50 μm. CD31 and Tie2 signals gradually increased in blood vessels, whereas CD34, CD146, and agrin signals gradually decreased. (B) The graphs show temporal changes in the percentage of marker+ area relative to the total vessel area. The colocalized signals between the markers and blood vessels were calculated, and the areas of the colocalized signals were normalized to the total blood vessel area in each image. Approximately 9–15 Images were obtained from the cerebral cortical regions of three rat pups of each age with a Nikon A1R-A1 confocal microscope. Data are shown as averaged value ± SEM. Data were analyzed using an ANOVA followed by a Tukey’s multiple range test. * p < 0.05, ** p < 0.01. vs. P1 value, while # p < 0.05, ## p < 0.01 vs. P10 value. (C) Graph summarizing results for B and B. Changes in biotin permeability, CD34, CD146, and agrin were corrected to a P1 value of 100%, and the astrocyte coverage rate, microglial coverage rate, CD31, and Tie2 were corrected to a P30 value of 100%. The grey zone represents the organic formation term (P4P15). Abbreviations: CD31, cluster of differentiation 31 or platelet endothelial cell adhesion molecule 1; CD146, melanoma cell adhesion molecule; DAPI: 4′,6-diamidino-2-phenylindole; P, postnatal day; Tie2, TEK receptor tyrosine kinase; ANOVA, analysis of variance.

    Techniques Used: Expressing, Staining, Marker, Microscopy, Permeability

    Analysis of vascular TJ protein expression in the postnatal cerebral cortex. (A) Images of claudin-5, occludin, and ZO-1 co-stained with lectin and DAPI in the rat cerebral cortex at P1–30. Scale bar indicates 100 μm. Claudin-5 and occludin signals increased in blood vessels during development, whereas ZO-1 signals remained unchanged. (B) The graphs show changes in the percentage of marker+ area of the total vessel area at P4, P15, and P30. The colocalized signals between the markers and blood vessels were calculated, and the areas of the colocalized signals were normalized to the total blood vessel area in each image. Approximately 9–15 images were obtained from the cerebral cortical regions of three rat pups from each age group using a Nikon A1R-A1 confocal microscope. Data are shown as averaged value ± SEM. Data were analyzed using an ANOVA followed by a Tukey’s multiple range test. * p < 0.05, ** p < 0.01 vs. P4 value, and # p < 0.05 vs. P15 value. DAPI: 4′,6-diamidino-2-phenylindole; P, postnatal day; TJ, tight junction; ZO-1: zonula occludens-1; ANOVA, analysis of variance.
    Figure Legend Snippet: Analysis of vascular TJ protein expression in the postnatal cerebral cortex. (A) Images of claudin-5, occludin, and ZO-1 co-stained with lectin and DAPI in the rat cerebral cortex at P1–30. Scale bar indicates 100 μm. Claudin-5 and occludin signals increased in blood vessels during development, whereas ZO-1 signals remained unchanged. (B) The graphs show changes in the percentage of marker+ area of the total vessel area at P4, P15, and P30. The colocalized signals between the markers and blood vessels were calculated, and the areas of the colocalized signals were normalized to the total blood vessel area in each image. Approximately 9–15 images were obtained from the cerebral cortical regions of three rat pups from each age group using a Nikon A1R-A1 confocal microscope. Data are shown as averaged value ± SEM. Data were analyzed using an ANOVA followed by a Tukey’s multiple range test. * p < 0.05, ** p < 0.01 vs. P4 value, and # p < 0.05 vs. P15 value. DAPI: 4′,6-diamidino-2-phenylindole; P, postnatal day; TJ, tight junction; ZO-1: zonula occludens-1; ANOVA, analysis of variance.

    Techniques Used: Expressing, Staining, Marker, Microscopy

    Analysis of vascular expression of transporters and receptor proteins in the postnatal cerebral cortex. (A) Images of P-gp, BCRP, Glut1, and TfR co-stained with lectin and DAPI in the rat cerebral cortex at P1–30. Scale bar indicates 100 μm. The arrowhead shows the co-localization areas of the marker and lectin. P-gp and BCRP signals gradually increased in the blood vessels during development, whereas Glut1 and TfR signals were high in the blood vessels at P1 and remained unchanged. (B) The graphs show changes in the percentage of marker+ area of the total vessel area at P4, P15, and P30. The colocalized signals between the markers and blood vessels were calculated, and the areas of the colocalized signals were normalized to the total blood vessel area in each image. Approximately 9–15 images were obtained from the cerebral cortical regions of three rat pups from each age group using a Nikon A1R-A1 confocal microscope. Data are shown as averaged value ± SEM. Data were analyzed using an ANOVA followed by a Tukey’s multiple range test. * p < 0.05, ** p < 0.01 vs. P4 value. Abbreviations: BCRP: breast cancer resistance protein; DAPI: 4′,6-diamidino-2-phenylindole; Glut1: glucose transporter type 1; P: postnatal day; P-gp: P-glycoprotein; TfR: transferrin receptor; ANOVA, analysis of variance.
    Figure Legend Snippet: Analysis of vascular expression of transporters and receptor proteins in the postnatal cerebral cortex. (A) Images of P-gp, BCRP, Glut1, and TfR co-stained with lectin and DAPI in the rat cerebral cortex at P1–30. Scale bar indicates 100 μm. The arrowhead shows the co-localization areas of the marker and lectin. P-gp and BCRP signals gradually increased in the blood vessels during development, whereas Glut1 and TfR signals were high in the blood vessels at P1 and remained unchanged. (B) The graphs show changes in the percentage of marker+ area of the total vessel area at P4, P15, and P30. The colocalized signals between the markers and blood vessels were calculated, and the areas of the colocalized signals were normalized to the total blood vessel area in each image. Approximately 9–15 images were obtained from the cerebral cortical regions of three rat pups from each age group using a Nikon A1R-A1 confocal microscope. Data are shown as averaged value ± SEM. Data were analyzed using an ANOVA followed by a Tukey’s multiple range test. * p < 0.05, ** p < 0.01 vs. P4 value. Abbreviations: BCRP: breast cancer resistance protein; DAPI: 4′,6-diamidino-2-phenylindole; Glut1: glucose transporter type 1; P: postnatal day; P-gp: P-glycoprotein; TfR: transferrin receptor; ANOVA, analysis of variance.

    Techniques Used: Expressing, Staining, Marker, Microscopy

    Related Articles

    Incubation:

    Article Title: Search for marker proteins to assess blood–brain barrier development
    Article Snippet: .. Thereafter, the sections were rinsed and incubated for 1 h at room temperature in a solution containing lectin (1:200; DL1177; Vector Labs) and DAPI (1:1000; 342–07431; Dojindo). .. After a final round of rinsing, drying, and embedding in VectaShield (H-1000; Vector Labs), the stained sections were analyzed using a Nikon A1R-A1 confocal microscope.

    Article Title: Duvelisib upregulates p27 expression and leads to intestinal damage via the NEDD4L/CK1ε axis.
    Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.

    Article Title: Search for marker proteins to assess blood-brain barrier development.
    Article Snippet: .. Thereafter, the sections were rinsed and incubated for 1 h at room temperature in a solution containing lectin (1:200; DL1177; Vector Labs) and DAPI (1:1000; 342–07431; Dojindo). .. After a final round of rinsing, drying, and embedding in VectaShield (H-1000; Vector Labs), the stained sections were analyzed using a Nikon A1R-A1 confocal microscope.

    Staining:

    Article Title: Duvelisib upregulates p27 expression and leads to intestinal damage via the NEDD4L/CK1ε axis.
    Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.

    Article Title: Search for marker proteins to assess blood-brain barrier development.
    Article Snippet: Subsequently, 30 μm thick sagittal sections of the embedded brains were cut, incubated at 24 °C for 3 h in a blocking solution (3% normal goat serum and 0.3% Triton X-100 in phosphate-buffered saline [PBS]), followed by incubation for 1 h at room temperature in a solution containing Alexa 546 streptavidin (1:500; S11225, Invitrogen, Waltham, MA, USA). .. After rinsing, the sections were stained with lectin (1:200; DL1177; Vector Labs, Newark, CA, USA) and 4′,6-diamidino-2-phenylindole (DAPI; 1:1000; 342-07431, Dojindo, Kumamoto, Japan). .. The stained sections were analyzed using a Nikon A1R confocal microscope system (Nikon, Tokyo, Japan) and quantified using NIS-Element analysis software (Nikon).

    Article Title: Search for marker proteins to assess blood–brain barrier development
    Article Snippet: Subsequently, 30 μm thick sagittal sections of the embedded brains were cut, incubated at 24 °C for 3 h in a blocking solution (3% normal goat serum and 0.3% Triton X-100 in phosphate-buffered saline [PBS]), followed by incubation for 1 h at room temperature in a solution containing Alexa 546 streptavidin (1:500; S11225 , Invitrogen, Waltham, MA, USA). .. After rinsing, the sections were stained with lectin (1:200; DL1177; Vector Labs, Newark, CA, USA) and 4′,6-diamidino-2-phenylindole (DAPI; 1:1000; 342-07431, Dojindo, Kumamoto, Japan). .. The stained sections were analyzed using a Nikon A1R confocal microscope system (Nikon, Tokyo, Japan) and quantified using NIS-Element analysis software (Nikon).

    Article Title: B7-H3 interaction with cMet enhances lipolysis to drive malignant progression of glioma via pSTAT3-ATGL axis.
    Article Snippet: .. Finally, DAPI (#D212, Dojindo Laboratories, Kumamoto, Japan) was then used for staining the nuclei, and images were acquired with Pannoramic MIDI II (3DHISTECH Ltd). .. For Lipid droplets visualization, the fixed glioma cells were rinsed 3 times with PBS and stained with BODIPY 493/503 (#D3922, Invitrogen) for 30 min at RT.

    Fluorescence:

    Article Title: Duvelisib upregulates p27 expression and leads to intestinal damage via the NEDD4L/CK1ε axis.
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    Microscopy:

    Article Title: Duvelisib upregulates p27 expression and leads to intestinal damage via the NEDD4L/CK1ε axis.
    Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.



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