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cy5 azide  (Jena Bioscience)


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    Structured Review

    Jena Bioscience cy5 azide
    Cy5 Azide, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 94/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cy5-azide/Cy5-Azide/pmc12876308-65-30-32
    Average 94 stars, based on 16 article reviews
    cy5 azide - by Bioz Stars, 2026-10
    94/100 stars

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    Incubation:

    Article Title: Isolation of proteins on chromatin (iPOC) reveals signaling pathway-dependent alterations in the DNA-bound proteome.
    Article Snippet: min. Cells were then spun down for 5 min at 900g and washed once with cold 0.5% BSA in 1x PBS, and once in 1x PBS. Permeabilized cells were then resuspend in click reaction buffer (10 μM biotin azide (Jena Bioscience CLK-047), 10 mM sodium ascorbate, 2 mM CuSO4 in PBS 1%) at RT, in the dark for 2h. Cells were then spun down for 5 min at 900g, washed once with cold 0.5% BSA in PBS, and once with 1x

    Lysis:

    Article Title: Isolation of proteins on chromatin (iPOC) reveals signaling pathway-dependent alterations in the DNA-bound proteome.
    Article Snippet: min. Cells were then spun down for 5 min at 900g and washed once with cold 0.5% BSA in 1x PBS, and once in 1x PBS. Permeabilized cells were then resuspend in click reaction buffer (10 μM biotin azide (Jena Bioscience CLK-047), 10 mM sodium ascorbate, 2 mM CuSO4 in PBS 1%) at RT, in the dark for 2h. Cells were then spun down for 5 min at 900g, washed once with cold 0.5% BSA in PBS, and once with 1x



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    Preparation and characterization of RGD‐EV:MiR‐9‐5p. (A) Western blot was used to detect the expression of the positive markers Alix and TSG101 in EVs. (B) Representative images of TTC‐stained brains from I/R model mice at 24 h after tail vein injection of PBS or EVs. (C) Quantification of infarct volume after TTC staining, n = 3. (D) Neurological deficit scores in the sham group, and I/R model mice treated with EVs or PBS. (E) RT‐PCR analysis of the levels of 10 different miRNAs in HEK293T‐derived EVs and ReN‐derived EVs ( n = 3). (F) After EV ReN was added to HT‐22 cells for 24 h, OGD experiments were performed and 5 miRNAs were detected ( n = 3). *denotes comparison with the Control group, *** p < 0.001; # denotes comparison between the OGD group and the OGD + EV ReN group, ### p < 0.001. (G) The binding sites of miR‐9‐5p on Hdac5 were predicted by TargetScan. (H) Dual‐luciferase reporter gene assay was used to verify the binding between miR‐9‐5p and Hdac5 . (I) BV2 cells were transfected with miR‐9‐5p mimics, and after 48 h, the expression of HDAC5 in the cells was analyzed by Western blot. (J) Quantification of HDAC5 ( n = 4). (K) Schematic of RGD‐EV, labeled with <t>Cy5.5,</t> and loaded with cholesterol‐modified miR‐9‐5p. (L, M) TEM imaging (L) and NTA (M) of RGD‐EV: MiR‐9‐5p; scale bar 200 nm. (N, O) MCAO/R mice were injected with Cy5.5‐labeled RGD‐EVs or RGD‐EV: MiR‐9‐5p, and Cy5.5 levels in the brain were analyzed 24 h later. Representative brain NIR fluorescence images (N) and quantification of intracerebral fluorescence intensity (O) are shown, n = 3. (P) After ischemia modeling in mice, tdTomato‐labeled EVs were injected into the tail vein and perfused 24 h later. Fluorescent staining was performed to analyze whether EVs had entered into microglia in the ischemic region; red indicates EVs, green indicates microglia, and the scale bar is 25 μm. (Q, R) BV2 cells were pretreated with PBS, EV (or EV: MiR‐NC, or EV: MiR‐9‐5p) for 24 h before LPS treatment. The expression of HDAC5 (Q) in the cells were determined by Western blot analysis, and quantification of HDAC5 (R), n = 3. Data are expressed as mean ± SEM, * p < 0.05; ** p < 0.01; *** p < 0.001 by one‐way ANOVA.
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    Preparation and characterization of RGD‐EV:MiR‐9‐5p. (A) Western blot was used to detect the expression of the positive markers Alix and TSG101 in EVs. (B) Representative images of TTC‐stained brains from I/R model mice at 24 h after tail vein injection of PBS or EVs. (C) Quantification of infarct volume after TTC staining, n = 3. (D) Neurological deficit scores in the sham group, and I/R model mice treated with EVs or PBS. (E) RT‐PCR analysis of the levels of 10 different miRNAs in HEK293T‐derived EVs and ReN‐derived EVs ( n = 3). (F) After EV ReN was added to HT‐22 cells for 24 h, OGD experiments were performed and 5 miRNAs were detected ( n = 3). *denotes comparison with the Control group, *** p < 0.001; # denotes comparison between the OGD group and the OGD + EV ReN group, ### p < 0.001. (G) The binding sites of miR‐9‐5p on Hdac5 were predicted by TargetScan. (H) Dual‐luciferase reporter gene assay was used to verify the binding between miR‐9‐5p and Hdac5 . (I) BV2 cells were transfected with miR‐9‐5p mimics, and after 48 h, the expression of HDAC5 in the cells was analyzed by Western blot. (J) Quantification of HDAC5 ( n = 4). (K) Schematic of RGD‐EV, labeled with <t>Cy5.5,</t> and loaded with cholesterol‐modified miR‐9‐5p. (L, M) TEM imaging (L) and NTA (M) of RGD‐EV: MiR‐9‐5p; scale bar 200 nm. (N, O) MCAO/R mice were injected with Cy5.5‐labeled RGD‐EVs or RGD‐EV: MiR‐9‐5p, and Cy5.5 levels in the brain were analyzed 24 h later. Representative brain NIR fluorescence images (N) and quantification of intracerebral fluorescence intensity (O) are shown, n = 3. (P) After ischemia modeling in mice, tdTomato‐labeled EVs were injected into the tail vein and perfused 24 h later. Fluorescent staining was performed to analyze whether EVs had entered into microglia in the ischemic region; red indicates EVs, green indicates microglia, and the scale bar is 25 μm. (Q, R) BV2 cells were pretreated with PBS, EV (or EV: MiR‐NC, or EV: MiR‐9‐5p) for 24 h before LPS treatment. The expression of HDAC5 (Q) in the cells were determined by Western blot analysis, and quantification of HDAC5 (R), n = 3. Data are expressed as mean ± SEM, * p < 0.05; ** p < 0.01; *** p < 0.001 by one‐way ANOVA.
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    Image Search Results


    Preparation and characterization of RGD‐EV:MiR‐9‐5p. (A) Western blot was used to detect the expression of the positive markers Alix and TSG101 in EVs. (B) Representative images of TTC‐stained brains from I/R model mice at 24 h after tail vein injection of PBS or EVs. (C) Quantification of infarct volume after TTC staining, n = 3. (D) Neurological deficit scores in the sham group, and I/R model mice treated with EVs or PBS. (E) RT‐PCR analysis of the levels of 10 different miRNAs in HEK293T‐derived EVs and ReN‐derived EVs ( n = 3). (F) After EV ReN was added to HT‐22 cells for 24 h, OGD experiments were performed and 5 miRNAs were detected ( n = 3). *denotes comparison with the Control group, *** p < 0.001; # denotes comparison between the OGD group and the OGD + EV ReN group, ### p < 0.001. (G) The binding sites of miR‐9‐5p on Hdac5 were predicted by TargetScan. (H) Dual‐luciferase reporter gene assay was used to verify the binding between miR‐9‐5p and Hdac5 . (I) BV2 cells were transfected with miR‐9‐5p mimics, and after 48 h, the expression of HDAC5 in the cells was analyzed by Western blot. (J) Quantification of HDAC5 ( n = 4). (K) Schematic of RGD‐EV, labeled with Cy5.5, and loaded with cholesterol‐modified miR‐9‐5p. (L, M) TEM imaging (L) and NTA (M) of RGD‐EV: MiR‐9‐5p; scale bar 200 nm. (N, O) MCAO/R mice were injected with Cy5.5‐labeled RGD‐EVs or RGD‐EV: MiR‐9‐5p, and Cy5.5 levels in the brain were analyzed 24 h later. Representative brain NIR fluorescence images (N) and quantification of intracerebral fluorescence intensity (O) are shown, n = 3. (P) After ischemia modeling in mice, tdTomato‐labeled EVs were injected into the tail vein and perfused 24 h later. Fluorescent staining was performed to analyze whether EVs had entered into microglia in the ischemic region; red indicates EVs, green indicates microglia, and the scale bar is 25 μm. (Q, R) BV2 cells were pretreated with PBS, EV (or EV: MiR‐NC, or EV: MiR‐9‐5p) for 24 h before LPS treatment. The expression of HDAC5 (Q) in the cells were determined by Western blot analysis, and quantification of HDAC5 (R), n = 3. Data are expressed as mean ± SEM, * p < 0.05; ** p < 0.01; *** p < 0.001 by one‐way ANOVA.

    Journal: CNS Neuroscience & Therapeutics

    Article Title: Targeting Microglial HDAC5 Suppresses NLRP3 Inflammation via NF ‐ κB Deacetylation to Alleviate Ischemic Brain Injury

    doi: 10.1002/cns.71161

    Figure Lengend Snippet: Preparation and characterization of RGD‐EV:MiR‐9‐5p. (A) Western blot was used to detect the expression of the positive markers Alix and TSG101 in EVs. (B) Representative images of TTC‐stained brains from I/R model mice at 24 h after tail vein injection of PBS or EVs. (C) Quantification of infarct volume after TTC staining, n = 3. (D) Neurological deficit scores in the sham group, and I/R model mice treated with EVs or PBS. (E) RT‐PCR analysis of the levels of 10 different miRNAs in HEK293T‐derived EVs and ReN‐derived EVs ( n = 3). (F) After EV ReN was added to HT‐22 cells for 24 h, OGD experiments were performed and 5 miRNAs were detected ( n = 3). *denotes comparison with the Control group, *** p < 0.001; # denotes comparison between the OGD group and the OGD + EV ReN group, ### p < 0.001. (G) The binding sites of miR‐9‐5p on Hdac5 were predicted by TargetScan. (H) Dual‐luciferase reporter gene assay was used to verify the binding between miR‐9‐5p and Hdac5 . (I) BV2 cells were transfected with miR‐9‐5p mimics, and after 48 h, the expression of HDAC5 in the cells was analyzed by Western blot. (J) Quantification of HDAC5 ( n = 4). (K) Schematic of RGD‐EV, labeled with Cy5.5, and loaded with cholesterol‐modified miR‐9‐5p. (L, M) TEM imaging (L) and NTA (M) of RGD‐EV: MiR‐9‐5p; scale bar 200 nm. (N, O) MCAO/R mice were injected with Cy5.5‐labeled RGD‐EVs or RGD‐EV: MiR‐9‐5p, and Cy5.5 levels in the brain were analyzed 24 h later. Representative brain NIR fluorescence images (N) and quantification of intracerebral fluorescence intensity (O) are shown, n = 3. (P) After ischemia modeling in mice, tdTomato‐labeled EVs were injected into the tail vein and perfused 24 h later. Fluorescent staining was performed to analyze whether EVs had entered into microglia in the ischemic region; red indicates EVs, green indicates microglia, and the scale bar is 25 μm. (Q, R) BV2 cells were pretreated with PBS, EV (or EV: MiR‐NC, or EV: MiR‐9‐5p) for 24 h before LPS treatment. The expression of HDAC5 (Q) in the cells were determined by Western blot analysis, and quantification of HDAC5 (R), n = 3. Data are expressed as mean ± SEM, * p < 0.05; ** p < 0.01; *** p < 0.001 by one‐way ANOVA.

    Article Snippet: For imaging, Cy5.5 azide (0.3 nmol; Lumiprobe, USA) was added simultaneously.

    Techniques: Western Blot, Expressing, Staining, Injection, Reverse Transcription Polymerase Chain Reaction, Analysis, Derivative Assay, Comparison, Control, Binding Assay, Luciferase, Reporter Gene Assay, Transfection, Labeling, Modification, Imaging, Fluorescence