vector blue alkaline phosphatase substrate kit iii  (Vector Laboratories)


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    BCIP NBT Alkaline Phosphatase AP Substrate Kit 5 bromo 4 chloro 3 indolyl phosphate nitroblue tetrazolium
    Description:
    BCIP NBT Alkaline Phosphatase Substrate Kit has greater sensitivity than conventional substratesConsistent and reliableIdeal for IHC ICC ISH and blotsFluorescentHeat StablePermanent mountingIdeal for single and multiple labelingOne year expiry dateStock solutions supplied in convenient dropper bottles promoting ease of handlingNo wait times for mixing and dissolving powders or tabletsSufficient reagents to produce 200 ml of working solutionExcitation 645 685 nm Emission peaks at 823 and 855 nm Biotechniques 2007 42 756 759 and BioTechniques 2014 57 254 256 BCIP NBT AP Substrate Kit forms an indigo heat stable reaction product that can be dehydrated cleared and permanently mounted This chromogen can be used singly or in combination with other alkaline phosphatase or peroxidase substrates for multiple label applications This substrate can also be used for developing blots or for in situ hybridization At 20 30 minute development times the sensitivity of this substrate is equivalent to the other alkaline phosphatase substrates However BCIP NBT will continue to develop from several hours to overnight making it one of the most sensitive chromogenic substrates This kit contains stock solutions in convenient dropper bottles
    Catalog Number:
    SK-5400
    Price:
    None
    Category:
    Protein chromogenic detection reagents or kits or substrates
    Size:
    1 Kit
    Buy from Supplier


    Structured Review

    Vector Laboratories vector blue alkaline phosphatase substrate kit iii
    BCIP NBT Alkaline Phosphatase AP Substrate Kit 5 bromo 4 chloro 3 indolyl phosphate nitroblue tetrazolium
    BCIP NBT Alkaline Phosphatase Substrate Kit has greater sensitivity than conventional substratesConsistent and reliableIdeal for IHC ICC ISH and blotsFluorescentHeat StablePermanent mountingIdeal for single and multiple labelingOne year expiry dateStock solutions supplied in convenient dropper bottles promoting ease of handlingNo wait times for mixing and dissolving powders or tabletsSufficient reagents to produce 200 ml of working solutionExcitation 645 685 nm Emission peaks at 823 and 855 nm Biotechniques 2007 42 756 759 and BioTechniques 2014 57 254 256 BCIP NBT AP Substrate Kit forms an indigo heat stable reaction product that can be dehydrated cleared and permanently mounted This chromogen can be used singly or in combination with other alkaline phosphatase or peroxidase substrates for multiple label applications This substrate can also be used for developing blots or for in situ hybridization At 20 30 minute development times the sensitivity of this substrate is equivalent to the other alkaline phosphatase substrates However BCIP NBT will continue to develop from several hours to overnight making it one of the most sensitive chromogenic substrates This kit contains stock solutions in convenient dropper bottles
    https://www.bioz.com/result/vector blue alkaline phosphatase substrate kit iii/product/Vector Laboratories
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vector blue alkaline phosphatase substrate kit iii - by Bioz Stars, 2021-05
    95/100 stars

    Images

    1) Product Images from "Ash2l interacts with Oct4-stemness circuitry to promote super-enhancer-driven pluripotency network"

    Article Title: Ash2l interacts with Oct4-stemness circuitry to promote super-enhancer-driven pluripotency network

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkz801

    Ash2l binding motif is crucial for the recruitment of OSN to Jarid2, Oct4 and Nanog super-enhancers. ( A ) Schematic illustration of the design of sequence-specific sgRNA for blocking of Jarid2, Nanog or Oct4 super-enhancers using monoallelic silencing (labeled as sgRNA AB ) or biallelic silencing (labeled as sgRNA AC , sgRNA DE , sgRNA DF , sgRNA GH and sgRNA GI ). The dCas9 nuclease is targeted to super-enhancer of Jarid2, Nanog or Oct4 by either set of sgRNA. Three amplicons for each enhancer (Jarid2: R1: enhancer; R2: promoter, R3: desert; Nanog: R4: enhancer; R5: promoter, R6: desert; Oct4: R7: enhancer; R8: promoter, R9: desert) were designed to evaluate protein enrichment in ChIP-qPCR assay. ( B ) ChIP-qPCR analysis showing the enrichment of indicated protein at Jarid2, Nanog and Oct4 super-enhancers in ESCs with CRISPRi/dCas9-mediated interference of Ash2l binding. ChIP-qPCR analysis with indicated antibodies was conducted in CRISPRi/dCas9-modified ESCs, and the enrichment of each protein at R1–3 regions, R4–6 regions or R7-R9 regions was evaluated with specific primers. Data showed that CRISPRi/dCas9-mediated interference of Ash2l binding at Jarid2 enhancer (upper), Nanog enhancer (middle) and Oct4 enhancer (lower), specifically increased dCas9 binding and decreased the binding of Ash2l, Oct4, Sox2, Nanog and H3K27ac, to R1, R4 and R7, but not R2, R3, R5, R6, R8 and R9. ( C ) qPCR results showing the effect of CRISPRi/dCas9-mediated interference of Ash2l binding on mRNA expression of Jarid2, Nanog and Oct4. ( D ) Western blot showing the effect of CRISPRi/dCas9-mediated interference of Ash2l binding on the protein content of Jarid2, Nanog and Oct4. ( E ) qPCR shows CRISPRi/dCas9-mediated interference of Ash2l binding at indicated enhancers abrogate the upregulation of Jarid2, Nanog, Oct4 transcripts induced by Ash2l overexpression. ( F ) Reporter assay shows that Flag-Ash2l and Myc-Oct4 induce a synergistic effect on enhancer activity in Jarid2, Nanog and Oct4 super-enhancers. The Ash2l-mediated enhancement of Jarid2, Nanog and Oct4 enhancer activation can be abrogated by specific sgRNA for indicated enhancer. Data are presented as mean ± SD; *, P
    Figure Legend Snippet: Ash2l binding motif is crucial for the recruitment of OSN to Jarid2, Oct4 and Nanog super-enhancers. ( A ) Schematic illustration of the design of sequence-specific sgRNA for blocking of Jarid2, Nanog or Oct4 super-enhancers using monoallelic silencing (labeled as sgRNA AB ) or biallelic silencing (labeled as sgRNA AC , sgRNA DE , sgRNA DF , sgRNA GH and sgRNA GI ). The dCas9 nuclease is targeted to super-enhancer of Jarid2, Nanog or Oct4 by either set of sgRNA. Three amplicons for each enhancer (Jarid2: R1: enhancer; R2: promoter, R3: desert; Nanog: R4: enhancer; R5: promoter, R6: desert; Oct4: R7: enhancer; R8: promoter, R9: desert) were designed to evaluate protein enrichment in ChIP-qPCR assay. ( B ) ChIP-qPCR analysis showing the enrichment of indicated protein at Jarid2, Nanog and Oct4 super-enhancers in ESCs with CRISPRi/dCas9-mediated interference of Ash2l binding. ChIP-qPCR analysis with indicated antibodies was conducted in CRISPRi/dCas9-modified ESCs, and the enrichment of each protein at R1–3 regions, R4–6 regions or R7-R9 regions was evaluated with specific primers. Data showed that CRISPRi/dCas9-mediated interference of Ash2l binding at Jarid2 enhancer (upper), Nanog enhancer (middle) and Oct4 enhancer (lower), specifically increased dCas9 binding and decreased the binding of Ash2l, Oct4, Sox2, Nanog and H3K27ac, to R1, R4 and R7, but not R2, R3, R5, R6, R8 and R9. ( C ) qPCR results showing the effect of CRISPRi/dCas9-mediated interference of Ash2l binding on mRNA expression of Jarid2, Nanog and Oct4. ( D ) Western blot showing the effect of CRISPRi/dCas9-mediated interference of Ash2l binding on the protein content of Jarid2, Nanog and Oct4. ( E ) qPCR shows CRISPRi/dCas9-mediated interference of Ash2l binding at indicated enhancers abrogate the upregulation of Jarid2, Nanog, Oct4 transcripts induced by Ash2l overexpression. ( F ) Reporter assay shows that Flag-Ash2l and Myc-Oct4 induce a synergistic effect on enhancer activity in Jarid2, Nanog and Oct4 super-enhancers. The Ash2l-mediated enhancement of Jarid2, Nanog and Oct4 enhancer activation can be abrogated by specific sgRNA for indicated enhancer. Data are presented as mean ± SD; *, P

    Techniques Used: Binding Assay, Sequencing, Blocking Assay, Labeling, Protein Enrichment, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Modification, Expressing, Western Blot, Over Expression, Reporter Assay, Activity Assay, Activation Assay

    2) Product Images from "Heme–hemopexin complex attenuates neuronal cell death and stroke damage"

    Article Title: Heme–hemopexin complex attenuates neuronal cell death and stroke damage

    Journal: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism

    doi: 10.1038/jcbfm.2009.19

    Heme–HPX induces HO1 expression in primary cortical neurons. Embryonic cortical neurons were incubated for 6 h with HHPX. HO1 induction was measured by RT-PCR ( A ) to analyze the relative level of HO1 mRNA or by Western blot ( B ) to determine HO1 protein, both compared with actin as an internal loading control. Lane a: untreated; lanes b to e: 1, 5, 10, and 20 μ mol/L HHPX, respectively. The fold changes in HO1 (relative to actin) are means of three independent experiments. ** P
    Figure Legend Snippet: Heme–HPX induces HO1 expression in primary cortical neurons. Embryonic cortical neurons were incubated for 6 h with HHPX. HO1 induction was measured by RT-PCR ( A ) to analyze the relative level of HO1 mRNA or by Western blot ( B ) to determine HO1 protein, both compared with actin as an internal loading control. Lane a: untreated; lanes b to e: 1, 5, 10, and 20 μ mol/L HHPX, respectively. The fold changes in HO1 (relative to actin) are means of three independent experiments. ** P

    Techniques Used: Expressing, Incubation, Reverse Transcription Polymerase Chain Reaction, Western Blot

    3) Product Images from "Nasal delivery of H5N1 avian influenza vaccine formulated with GenJet™ or in vivo-jetPEI® induces enhanced serological, cellular and protective immune responses"

    Article Title: Nasal delivery of H5N1 avian influenza vaccine formulated with GenJet™ or in vivo-jetPEI® induces enhanced serological, cellular and protective immune responses

    Journal: Drug Delivery

    doi: 10.1080/10717544.2018.1450909

    GenJet™ and in vivo -jetPEI ® enhanced the H5N1-specific T-cell responses. Balb/c mice (3–5 mice/group) were intranasally administered with A/IN/05 vaccine with or without GenJet™ or in vivo -jetPEI ® using prime-boost regimen as described in Figure 1 . (A) One week after booster immunization, the lung tissues were harvested and single cell suspensions were prepared. About 10 6 cells from the lung were stimulated in vitro with HA peptide for 6 h to examine the antigen-specific CD8 T-cell response; or with RG A/IN/05 virus at an MOI of 1 for 16 h to examine the antigen-specific CD4 T-cell response. GolgiPlug™ was added during the last 5 h of incubation. Cells were surface stained with anti-CD44, anti-CD4 or anti-CD8 antibody (BD Bioscience), followed by intracellular staining with anti-IFNγ antibody (BD Bioscience). The frequency of IFN-γ producing T cells in total activated T cells was presented. (B) One-week post-booster immunization, the draining lymph nodes, lungs and spleen tissues were harvested and the frequency of HA518-specific CD8 T cells in total activated CD8 T cells was stained using H-2K d /IYSTVASSL tetramer. (C) Three weeks after booster immunization, sera were collected and IgG2a, IgG2b IgG3 and IgG1 antibodies against A/IN/05 were assessed by ELISA. The data are representative of two independent experiments (3–5 mice each group) and error bars represent SEM. One-way ANOVA with Bonferroni post-analysis was used to analyze differences among treatments. p
    Figure Legend Snippet: GenJet™ and in vivo -jetPEI ® enhanced the H5N1-specific T-cell responses. Balb/c mice (3–5 mice/group) were intranasally administered with A/IN/05 vaccine with or without GenJet™ or in vivo -jetPEI ® using prime-boost regimen as described in Figure 1 . (A) One week after booster immunization, the lung tissues were harvested and single cell suspensions were prepared. About 10 6 cells from the lung were stimulated in vitro with HA peptide for 6 h to examine the antigen-specific CD8 T-cell response; or with RG A/IN/05 virus at an MOI of 1 for 16 h to examine the antigen-specific CD4 T-cell response. GolgiPlug™ was added during the last 5 h of incubation. Cells were surface stained with anti-CD44, anti-CD4 or anti-CD8 antibody (BD Bioscience), followed by intracellular staining with anti-IFNγ antibody (BD Bioscience). The frequency of IFN-γ producing T cells in total activated T cells was presented. (B) One-week post-booster immunization, the draining lymph nodes, lungs and spleen tissues were harvested and the frequency of HA518-specific CD8 T cells in total activated CD8 T cells was stained using H-2K d /IYSTVASSL tetramer. (C) Three weeks after booster immunization, sera were collected and IgG2a, IgG2b IgG3 and IgG1 antibodies against A/IN/05 were assessed by ELISA. The data are representative of two independent experiments (3–5 mice each group) and error bars represent SEM. One-way ANOVA with Bonferroni post-analysis was used to analyze differences among treatments. p

    Techniques Used: In Vivo, Mouse Assay, In Vitro, Incubation, Staining, Enzyme-linked Immunosorbent Assay

    GenJet™ and in vivo -jetPEI enhances the H5N1 vaccine-induced systemic antibody responses and memory B-cell responses. Balb/c mice (5 mice/group) were intranasally administered with 3 µg of A/IN/05 vaccine with or without GenJet™ (10 µl/each mouse as described in previous study (Kulkarni et al., 2014 )) or in vivo -jetPEI ® (0.6 μl/mouse according to manufacturer’s protocol). One month later, mice were boosted with the same vaccine formulations. The control mouse group received GenJet™, in vivo -jetPEI ® or PBS at both time points. (A) Three weeks after booster immunization, sera were collected and IgG, IgA and IgM antibodies against A/IN/05 were assessed by ELISA. (B) One week after booster immunization, the spleen were harvested and the frequency of A/IN/05-specific IgG + ASCs in the spleen were measured by ELISPOT assay. The number of A/IN/05-specific IgG + ASCs were normalized against the number of total IgG + secreting ASCs and presented as % Ag-specific IgG + B cells. The data are representative of two independent experiments (3–5 mice each group) and error bars represent SEM. One-way ANOVA with Bonferroni post-analysis was used to analyze differences among different groups. p
    Figure Legend Snippet: GenJet™ and in vivo -jetPEI enhances the H5N1 vaccine-induced systemic antibody responses and memory B-cell responses. Balb/c mice (5 mice/group) were intranasally administered with 3 µg of A/IN/05 vaccine with or without GenJet™ (10 µl/each mouse as described in previous study (Kulkarni et al., 2014 )) or in vivo -jetPEI ® (0.6 μl/mouse according to manufacturer’s protocol). One month later, mice were boosted with the same vaccine formulations. The control mouse group received GenJet™, in vivo -jetPEI ® or PBS at both time points. (A) Three weeks after booster immunization, sera were collected and IgG, IgA and IgM antibodies against A/IN/05 were assessed by ELISA. (B) One week after booster immunization, the spleen were harvested and the frequency of A/IN/05-specific IgG + ASCs in the spleen were measured by ELISPOT assay. The number of A/IN/05-specific IgG + ASCs were normalized against the number of total IgG + secreting ASCs and presented as % Ag-specific IgG + B cells. The data are representative of two independent experiments (3–5 mice each group) and error bars represent SEM. One-way ANOVA with Bonferroni post-analysis was used to analyze differences among different groups. p

    Techniques Used: In Vivo, Mouse Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

    Related Articles

    Staining:

    Article Title: Small molecules facilitate rapid and synchronous iPSC generation
    Article Snippet: .. Alkaline phosphatase staining Alkaline phosphatase kits (Vector Labs) were used according to the manufacturer’s recommendations to assess pluripotency. .. Prior to staining, doxycycline and cell culture supplements were removed for a minimum of 3–4 days to eliminate exogenous OKSM expression.

    Article Title: Active induction of experimental autoimmune encephalomyelitis by MOG35-55 peptide immunization is associated with differential responses in separate compartments of the choroid plexus
    Article Snippet: Immunostaining for Immuno-LCM Immunostaining was performed as detailed [ , , ], with minor modifications. .. The CP stromal capillaries were stained using alkaline phosphatase substrate NBT (nitro-blue tetrazolium chloride)/BCIP (5-bromo-4-chloro-3'-indolyphosphate p-toluidine salt), (Vector Labs, Burlingame, CA) for 3–5 minutes in 100 Mm Tris–HCl (pH 9.5) to detect endogenous alkaline phosphatase activity in the endothelial cells. ..

    Article Title: Butyrophilin-like 1 encodes an enterocyte protein that selectively regulates functional interactions with T lymphocytes
    Article Snippet: Acetone-fixed frozen sections or Methanol-Carnoy's fixed and paraffin-embedded sections were stained with anti-Btnl1 only or double stained for expression of Btnl1 and CD3. .. The alkaline phosphatase conjugated anti-CD3 staining was developed with Vector Blue Alkaline Phosphatase Substrate Kit III (Vector Laboratories) and peroxidase stained Btnl1 sections developed using the 3, 3′-diaminobenzidine substrate kit (Vector Laboratories). .. Frozen sections or Methanol-Carnoy's fixed and paraffin embedded sections were double stained for Btnl1 and CD3 or Btnl1 and A33.

    Activity Assay:

    Article Title: Active induction of experimental autoimmune encephalomyelitis by MOG35-55 peptide immunization is associated with differential responses in separate compartments of the choroid plexus
    Article Snippet: Immunostaining for Immuno-LCM Immunostaining was performed as detailed [ , , ], with minor modifications. .. The CP stromal capillaries were stained using alkaline phosphatase substrate NBT (nitro-blue tetrazolium chloride)/BCIP (5-bromo-4-chloro-3'-indolyphosphate p-toluidine salt), (Vector Labs, Burlingame, CA) for 3–5 minutes in 100 Mm Tris–HCl (pH 9.5) to detect endogenous alkaline phosphatase activity in the endothelial cells. ..

    Plasmid Preparation:

    Article Title: Heme–hemopexin complex attenuates neuronal cell death and stroke damage
    Article Snippet: This antibody cross-reacts with mouse HPX and the secondary antibody was selected accordingly (Vector Laboratories). .. The HPX was finally visualized by developing the sections with the Vector Blue Alkaline Phosphatase Substrate Kit III (Vector Laboratories). .. Cultured embryonic neurons were grown on poly-D-lysine-coated glass coverslips for 10 days.

    Article Title: Butyrophilin-like 1 encodes an enterocyte protein that selectively regulates functional interactions with T lymphocytes
    Article Snippet: Acetone-fixed frozen sections or Methanol-Carnoy's fixed and paraffin-embedded sections were stained with anti-Btnl1 only or double stained for expression of Btnl1 and CD3. .. The alkaline phosphatase conjugated anti-CD3 staining was developed with Vector Blue Alkaline Phosphatase Substrate Kit III (Vector Laboratories) and peroxidase stained Btnl1 sections developed using the 3, 3′-diaminobenzidine substrate kit (Vector Laboratories). .. Frozen sections or Methanol-Carnoy's fixed and paraffin embedded sections were double stained for Btnl1 and CD3 or Btnl1 and A33.

    Article Title: RGC-32 regulates reactive astrocytosis and extracellular matrix deposition in experimental autoimmune encephalomyelitis
    Article Snippet: Frozen sections (5 μm) from the brains of adult patients with MS were double-stained for collagen I, IV, or V and for GFAP as previously described [ ]. .. Cryosections were initially treated with Bloxall (Vector Labs) to remove endogenous peroxidase and alkaline phosphatase, then incubated with primary antibodies against collagen I, IV, or V as described above, and the reaction was developed with Nova RED (Vector Labs) for collagen I and RGC-32 or Vector alkaline phosphatase substrate kit III (Vector Labs) for collagens IV and V. The sections were then incubated overnight with mouse IgG anti-GFAP monoclonal antibody (eBioscience, San Diego, CA). .. The slides were washed several times in PBS and reacted with alkaline phosphatase-conjugated goat anti-mouse (Sigma-Aldrich), diluted 1/400 and processed with Vector alkaline phosphatase substrate kit III (Vector Labs) or Nova RED as the chromogen substrate.

    Incubation:

    Article Title: Broad cross-reactive IgG responses elicited by adjuvanted vaccination with recombinant influenza hemagglutinin (rHA) in ferrets and mice
    Article Snippet: After the incubation, the wells were washed six times with 0.1% Tween-20 PBS (PBST, Sigma-Aldrich, St. Louis, MO) and phosphatase-labeled goat anti-mouse IgG detection antibody (catalog #1030–50, SouthernBiotech, AL) added at 100 μL/well. .. Plates were then incubated overnight at 4°C, washed six times with PBST, then IgG specific spots were developed using the Alkaline Phosphate Substrate Kit III (Vector Laboratories, Burlingame, Ca). .. Spots numbers were enumerated using an ImmunoSpot plate reader (Cellular Technology Limited, Cleveland, OH).

    Article Title: RGC-32 regulates reactive astrocytosis and extracellular matrix deposition in experimental autoimmune encephalomyelitis
    Article Snippet: Frozen sections (5 μm) from the brains of adult patients with MS were double-stained for collagen I, IV, or V and for GFAP as previously described [ ]. .. Cryosections were initially treated with Bloxall (Vector Labs) to remove endogenous peroxidase and alkaline phosphatase, then incubated with primary antibodies against collagen I, IV, or V as described above, and the reaction was developed with Nova RED (Vector Labs) for collagen I and RGC-32 or Vector alkaline phosphatase substrate kit III (Vector Labs) for collagens IV and V. The sections were then incubated overnight with mouse IgG anti-GFAP monoclonal antibody (eBioscience, San Diego, CA). .. The slides were washed several times in PBS and reacted with alkaline phosphatase-conjugated goat anti-mouse (Sigma-Aldrich), diluted 1/400 and processed with Vector alkaline phosphatase substrate kit III (Vector Labs) or Nova RED as the chromogen substrate.

    Article Title: Nuclear factor kappa B modulates apoptosis in the brain endothelial cells and intravascular leukocytes of fatal cerebral malaria
    Article Snippet: Sections were incubated overnight at 4°C with specific primary antibody; mouse monoclonal antibody directed against the p65 (F-6) RelA component of the NF-κB complex (1:50; Santa Cruz Biotechnology, CA, USA), and rabbit polyclonal antibody that recognized the large fragment (17/19 kDa) of cleaved caspase-3 (1:200; Cell Signaling Technology, USA). .. During the ensuing days, sections were washed three times with PBS and incubated with secondary antibody for 30 min at room temperature and reacted with avidin-biotin complex (ABC) conjugated with horseradish peroxidase (HRP) (for NF-κB p65) or alkaline phosphatase (AP) (for cleaved caspase-3) (Vectastain ABC kit, Vector Laboratories, CA, USA), according to the manufacturer’s instructions. .. After washing with PBS, enzymatic activity was visualized by 3,3'-diaminobenzidine tetrahydrochloride (DAB) or Vector® Red substrate kit (Vector Laboratories, CA, USA) for peroxidase (brown color) and alkaline phosphatase (red color) activity, respectively.

    Avidin-Biotin Assay:

    Article Title: Nuclear factor kappa B modulates apoptosis in the brain endothelial cells and intravascular leukocytes of fatal cerebral malaria
    Article Snippet: Sections were incubated overnight at 4°C with specific primary antibody; mouse monoclonal antibody directed against the p65 (F-6) RelA component of the NF-κB complex (1:50; Santa Cruz Biotechnology, CA, USA), and rabbit polyclonal antibody that recognized the large fragment (17/19 kDa) of cleaved caspase-3 (1:200; Cell Signaling Technology, USA). .. During the ensuing days, sections were washed three times with PBS and incubated with secondary antibody for 30 min at room temperature and reacted with avidin-biotin complex (ABC) conjugated with horseradish peroxidase (HRP) (for NF-κB p65) or alkaline phosphatase (AP) (for cleaved caspase-3) (Vectastain ABC kit, Vector Laboratories, CA, USA), according to the manufacturer’s instructions. .. After washing with PBS, enzymatic activity was visualized by 3,3'-diaminobenzidine tetrahydrochloride (DAB) or Vector® Red substrate kit (Vector Laboratories, CA, USA) for peroxidase (brown color) and alkaline phosphatase (red color) activity, respectively.

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    Vector Laboratories vector blue alkaline phosphatase substrate kit iii
    <t>Heme–HPX</t> induces HO1 expression in primary cortical neurons. Embryonic cortical neurons were incubated for 6 h with HHPX. HO1 induction was measured by RT-PCR ( A ) to analyze the relative level of HO1 mRNA or by Western blot ( B ) to determine HO1 protein, both compared with actin as an internal loading control. Lane a: untreated; lanes b to e: 1, 5, 10, and 20 μ mol/L HHPX, respectively. The fold changes in HO1 (relative to actin) are means of <t>three</t> independent experiments. ** P
    Vector Blue Alkaline Phosphatase Substrate Kit Iii, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vector blue alkaline phosphatase substrate kit iii/product/Vector Laboratories
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vector blue alkaline phosphatase substrate kit iii - by Bioz Stars, 2021-05
    95/100 stars
      Buy from Supplier

    Image Search Results


    Heme–HPX induces HO1 expression in primary cortical neurons. Embryonic cortical neurons were incubated for 6 h with HHPX. HO1 induction was measured by RT-PCR ( A ) to analyze the relative level of HO1 mRNA or by Western blot ( B ) to determine HO1 protein, both compared with actin as an internal loading control. Lane a: untreated; lanes b to e: 1, 5, 10, and 20 μ mol/L HHPX, respectively. The fold changes in HO1 (relative to actin) are means of three independent experiments. ** P

    Journal: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism

    Article Title: Heme–hemopexin complex attenuates neuronal cell death and stroke damage

    doi: 10.1038/jcbfm.2009.19

    Figure Lengend Snippet: Heme–HPX induces HO1 expression in primary cortical neurons. Embryonic cortical neurons were incubated for 6 h with HHPX. HO1 induction was measured by RT-PCR ( A ) to analyze the relative level of HO1 mRNA or by Western blot ( B ) to determine HO1 protein, both compared with actin as an internal loading control. Lane a: untreated; lanes b to e: 1, 5, 10, and 20 μ mol/L HHPX, respectively. The fold changes in HO1 (relative to actin) are means of three independent experiments. ** P

    Article Snippet: The HPX was finally visualized by developing the sections with the Vector Blue Alkaline Phosphatase Substrate Kit III (Vector Laboratories).

    Techniques: Expressing, Incubation, Reverse Transcription Polymerase Chain Reaction, Western Blot

    Quantitative RT-PCR analysis of Btnl1 , -4 , and -6 mRNA ( A ) in normal mouse tissues; ( B ) in CD45 (−) small intestinal epithelial cells (IECs) versus CD45 (+) small intestinal IEL; ( C ) in large intestines harvested from three mice; and ( D ) in the

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

    Article Title: Butyrophilin-like 1 encodes an enterocyte protein that selectively regulates functional interactions with T lymphocytes

    doi: 10.1073/pnas.1010647108

    Figure Lengend Snippet: Quantitative RT-PCR analysis of Btnl1 , -4 , and -6 mRNA ( A ) in normal mouse tissues; ( B ) in CD45 (−) small intestinal epithelial cells (IECs) versus CD45 (+) small intestinal IEL; ( C ) in large intestines harvested from three mice; and ( D ) in the

    Article Snippet: The alkaline phosphatase conjugated anti-CD3 staining was developed with Vector Blue Alkaline Phosphatase Substrate Kit III (Vector Laboratories) and peroxidase stained Btnl1 sections developed using the 3, 3′-diaminobenzidine substrate kit (Vector Laboratories).

    Techniques: Quantitative RT-PCR, Mouse Assay

    GenJet™ and in vivo -jetPEI ® enhanced the H5N1-specific T-cell responses. Balb/c mice (3–5 mice/group) were intranasally administered with A/IN/05 vaccine with or without GenJet™ or in vivo -jetPEI ® using prime-boost regimen as described in Figure 1 . (A) One week after booster immunization, the lung tissues were harvested and single cell suspensions were prepared. About 10 6 cells from the lung were stimulated in vitro with HA peptide for 6 h to examine the antigen-specific CD8 T-cell response; or with RG A/IN/05 virus at an MOI of 1 for 16 h to examine the antigen-specific CD4 T-cell response. GolgiPlug™ was added during the last 5 h of incubation. Cells were surface stained with anti-CD44, anti-CD4 or anti-CD8 antibody (BD Bioscience), followed by intracellular staining with anti-IFNγ antibody (BD Bioscience). The frequency of IFN-γ producing T cells in total activated T cells was presented. (B) One-week post-booster immunization, the draining lymph nodes, lungs and spleen tissues were harvested and the frequency of HA518-specific CD8 T cells in total activated CD8 T cells was stained using H-2K d /IYSTVASSL tetramer. (C) Three weeks after booster immunization, sera were collected and IgG2a, IgG2b IgG3 and IgG1 antibodies against A/IN/05 were assessed by ELISA. The data are representative of two independent experiments (3–5 mice each group) and error bars represent SEM. One-way ANOVA with Bonferroni post-analysis was used to analyze differences among treatments. p

    Journal: Drug Delivery

    Article Title: Nasal delivery of H5N1 avian influenza vaccine formulated with GenJet™ or in vivo-jetPEI® induces enhanced serological, cellular and protective immune responses

    doi: 10.1080/10717544.2018.1450909

    Figure Lengend Snippet: GenJet™ and in vivo -jetPEI ® enhanced the H5N1-specific T-cell responses. Balb/c mice (3–5 mice/group) were intranasally administered with A/IN/05 vaccine with or without GenJet™ or in vivo -jetPEI ® using prime-boost regimen as described in Figure 1 . (A) One week after booster immunization, the lung tissues were harvested and single cell suspensions were prepared. About 10 6 cells from the lung were stimulated in vitro with HA peptide for 6 h to examine the antigen-specific CD8 T-cell response; or with RG A/IN/05 virus at an MOI of 1 for 16 h to examine the antigen-specific CD4 T-cell response. GolgiPlug™ was added during the last 5 h of incubation. Cells were surface stained with anti-CD44, anti-CD4 or anti-CD8 antibody (BD Bioscience), followed by intracellular staining with anti-IFNγ antibody (BD Bioscience). The frequency of IFN-γ producing T cells in total activated T cells was presented. (B) One-week post-booster immunization, the draining lymph nodes, lungs and spleen tissues were harvested and the frequency of HA518-specific CD8 T cells in total activated CD8 T cells was stained using H-2K d /IYSTVASSL tetramer. (C) Three weeks after booster immunization, sera were collected and IgG2a, IgG2b IgG3 and IgG1 antibodies against A/IN/05 were assessed by ELISA. The data are representative of two independent experiments (3–5 mice each group) and error bars represent SEM. One-way ANOVA with Bonferroni post-analysis was used to analyze differences among treatments. p

    Article Snippet: The plates were incubated with biotinylated anti-mouse IgG (Southern Biotech) followed by alkaline phosphatase-conjugated streptavidin and developed with Vector Blue Alkaline Phosphatase Substrate Kit III (Vector Laboratories, Burlingame, CA).

    Techniques: In Vivo, Mouse Assay, In Vitro, Incubation, Staining, Enzyme-linked Immunosorbent Assay

    GenJet™ and in vivo -jetPEI enhances the H5N1 vaccine-induced systemic antibody responses and memory B-cell responses. Balb/c mice (5 mice/group) were intranasally administered with 3 µg of A/IN/05 vaccine with or without GenJet™ (10 µl/each mouse as described in previous study (Kulkarni et al., 2014 )) or in vivo -jetPEI ® (0.6 μl/mouse according to manufacturer’s protocol). One month later, mice were boosted with the same vaccine formulations. The control mouse group received GenJet™, in vivo -jetPEI ® or PBS at both time points. (A) Three weeks after booster immunization, sera were collected and IgG, IgA and IgM antibodies against A/IN/05 were assessed by ELISA. (B) One week after booster immunization, the spleen were harvested and the frequency of A/IN/05-specific IgG + ASCs in the spleen were measured by ELISPOT assay. The number of A/IN/05-specific IgG + ASCs were normalized against the number of total IgG + secreting ASCs and presented as % Ag-specific IgG + B cells. The data are representative of two independent experiments (3–5 mice each group) and error bars represent SEM. One-way ANOVA with Bonferroni post-analysis was used to analyze differences among different groups. p

    Journal: Drug Delivery

    Article Title: Nasal delivery of H5N1 avian influenza vaccine formulated with GenJet™ or in vivo-jetPEI® induces enhanced serological, cellular and protective immune responses

    doi: 10.1080/10717544.2018.1450909

    Figure Lengend Snippet: GenJet™ and in vivo -jetPEI enhances the H5N1 vaccine-induced systemic antibody responses and memory B-cell responses. Balb/c mice (5 mice/group) were intranasally administered with 3 µg of A/IN/05 vaccine with or without GenJet™ (10 µl/each mouse as described in previous study (Kulkarni et al., 2014 )) or in vivo -jetPEI ® (0.6 μl/mouse according to manufacturer’s protocol). One month later, mice were boosted with the same vaccine formulations. The control mouse group received GenJet™, in vivo -jetPEI ® or PBS at both time points. (A) Three weeks after booster immunization, sera were collected and IgG, IgA and IgM antibodies against A/IN/05 were assessed by ELISA. (B) One week after booster immunization, the spleen were harvested and the frequency of A/IN/05-specific IgG + ASCs in the spleen were measured by ELISPOT assay. The number of A/IN/05-specific IgG + ASCs were normalized against the number of total IgG + secreting ASCs and presented as % Ag-specific IgG + B cells. The data are representative of two independent experiments (3–5 mice each group) and error bars represent SEM. One-way ANOVA with Bonferroni post-analysis was used to analyze differences among different groups. p

    Article Snippet: The plates were incubated with biotinylated anti-mouse IgG (Southern Biotech) followed by alkaline phosphatase-conjugated streptavidin and developed with Vector Blue Alkaline Phosphatase Substrate Kit III (Vector Laboratories, Burlingame, CA).

    Techniques: In Vivo, Mouse Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

    Ash2l binding motif is crucial for the recruitment of OSN to Jarid2, Oct4 and Nanog super-enhancers. ( A ) Schematic illustration of the design of sequence-specific sgRNA for blocking of Jarid2, Nanog or Oct4 super-enhancers using monoallelic silencing (labeled as sgRNA AB ) or biallelic silencing (labeled as sgRNA AC , sgRNA DE , sgRNA DF , sgRNA GH and sgRNA GI ). The dCas9 nuclease is targeted to super-enhancer of Jarid2, Nanog or Oct4 by either set of sgRNA. Three amplicons for each enhancer (Jarid2: R1: enhancer; R2: promoter, R3: desert; Nanog: R4: enhancer; R5: promoter, R6: desert; Oct4: R7: enhancer; R8: promoter, R9: desert) were designed to evaluate protein enrichment in ChIP-qPCR assay. ( B ) ChIP-qPCR analysis showing the enrichment of indicated protein at Jarid2, Nanog and Oct4 super-enhancers in ESCs with CRISPRi/dCas9-mediated interference of Ash2l binding. ChIP-qPCR analysis with indicated antibodies was conducted in CRISPRi/dCas9-modified ESCs, and the enrichment of each protein at R1–3 regions, R4–6 regions or R7-R9 regions was evaluated with specific primers. Data showed that CRISPRi/dCas9-mediated interference of Ash2l binding at Jarid2 enhancer (upper), Nanog enhancer (middle) and Oct4 enhancer (lower), specifically increased dCas9 binding and decreased the binding of Ash2l, Oct4, Sox2, Nanog and H3K27ac, to R1, R4 and R7, but not R2, R3, R5, R6, R8 and R9. ( C ) qPCR results showing the effect of CRISPRi/dCas9-mediated interference of Ash2l binding on mRNA expression of Jarid2, Nanog and Oct4. ( D ) Western blot showing the effect of CRISPRi/dCas9-mediated interference of Ash2l binding on the protein content of Jarid2, Nanog and Oct4. ( E ) qPCR shows CRISPRi/dCas9-mediated interference of Ash2l binding at indicated enhancers abrogate the upregulation of Jarid2, Nanog, Oct4 transcripts induced by Ash2l overexpression. ( F ) Reporter assay shows that Flag-Ash2l and Myc-Oct4 induce a synergistic effect on enhancer activity in Jarid2, Nanog and Oct4 super-enhancers. The Ash2l-mediated enhancement of Jarid2, Nanog and Oct4 enhancer activation can be abrogated by specific sgRNA for indicated enhancer. Data are presented as mean ± SD; *, P

    Journal: Nucleic Acids Research

    Article Title: Ash2l interacts with Oct4-stemness circuitry to promote super-enhancer-driven pluripotency network

    doi: 10.1093/nar/gkz801

    Figure Lengend Snippet: Ash2l binding motif is crucial for the recruitment of OSN to Jarid2, Oct4 and Nanog super-enhancers. ( A ) Schematic illustration of the design of sequence-specific sgRNA for blocking of Jarid2, Nanog or Oct4 super-enhancers using monoallelic silencing (labeled as sgRNA AB ) or biallelic silencing (labeled as sgRNA AC , sgRNA DE , sgRNA DF , sgRNA GH and sgRNA GI ). The dCas9 nuclease is targeted to super-enhancer of Jarid2, Nanog or Oct4 by either set of sgRNA. Three amplicons for each enhancer (Jarid2: R1: enhancer; R2: promoter, R3: desert; Nanog: R4: enhancer; R5: promoter, R6: desert; Oct4: R7: enhancer; R8: promoter, R9: desert) were designed to evaluate protein enrichment in ChIP-qPCR assay. ( B ) ChIP-qPCR analysis showing the enrichment of indicated protein at Jarid2, Nanog and Oct4 super-enhancers in ESCs with CRISPRi/dCas9-mediated interference of Ash2l binding. ChIP-qPCR analysis with indicated antibodies was conducted in CRISPRi/dCas9-modified ESCs, and the enrichment of each protein at R1–3 regions, R4–6 regions or R7-R9 regions was evaluated with specific primers. Data showed that CRISPRi/dCas9-mediated interference of Ash2l binding at Jarid2 enhancer (upper), Nanog enhancer (middle) and Oct4 enhancer (lower), specifically increased dCas9 binding and decreased the binding of Ash2l, Oct4, Sox2, Nanog and H3K27ac, to R1, R4 and R7, but not R2, R3, R5, R6, R8 and R9. ( C ) qPCR results showing the effect of CRISPRi/dCas9-mediated interference of Ash2l binding on mRNA expression of Jarid2, Nanog and Oct4. ( D ) Western blot showing the effect of CRISPRi/dCas9-mediated interference of Ash2l binding on the protein content of Jarid2, Nanog and Oct4. ( E ) qPCR shows CRISPRi/dCas9-mediated interference of Ash2l binding at indicated enhancers abrogate the upregulation of Jarid2, Nanog, Oct4 transcripts induced by Ash2l overexpression. ( F ) Reporter assay shows that Flag-Ash2l and Myc-Oct4 induce a synergistic effect on enhancer activity in Jarid2, Nanog and Oct4 super-enhancers. The Ash2l-mediated enhancement of Jarid2, Nanog and Oct4 enhancer activation can be abrogated by specific sgRNA for indicated enhancer. Data are presented as mean ± SD; *, P

    Article Snippet: Alkaline phosphatase staining For detecting the alkaline phosphatase (AP) activity, reprogrammed MEFs, iPSCs or ESCs were fixed with 80% alcohol and then stained with the Vector Blue Alkaline Phosphatase Substrate Kit III (Vector Laboratories), according to the manufacturer’s instructions.

    Techniques: Binding Assay, Sequencing, Blocking Assay, Labeling, Protein Enrichment, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Modification, Expressing, Western Blot, Over Expression, Reporter Assay, Activity Assay, Activation Assay