streptavidin  (Vector Laboratories)


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    Name:
    Streptavidin Biotin Blocking Kit
    Description:
    Streptavidin Biotin Blocking Kit blocks all endogenous biotin biotin receptors and streptavidin binding sites present in tissues This kit is designed for use with streptavidin detection systems such as the VECTASTAIN Universal Quick kits if streptavidin or biotinylated reagents bind non specifically to tissues or proteins This blocking kit consists of 18 ml of streptavidin and 18 ml of biotin in convenient dropper bottles
    Catalog Number:
    sp-2002
    Price:
    None
    Category:
    Blocking agents
    Size:
    1 kit
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    Structured Review

    Vector Laboratories streptavidin
    Streptavidin Biotin Blocking Kit
    Streptavidin Biotin Blocking Kit blocks all endogenous biotin biotin receptors and streptavidin binding sites present in tissues This kit is designed for use with streptavidin detection systems such as the VECTASTAIN Universal Quick kits if streptavidin or biotinylated reagents bind non specifically to tissues or proteins This blocking kit consists of 18 ml of streptavidin and 18 ml of biotin in convenient dropper bottles
    https://www.bioz.com/result/streptavidin/product/Vector Laboratories
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin - by Bioz Stars, 2021-03
    94/100 stars

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    Related Articles

    Blocking Assay:

    Article Title: Antigen Display, T-Cell Activation, and Immune Evasion during Acute and Chronic Ehrlichiosis
    Article Snippet: For the detection of E. muris , biotinylated Ec18.1 (1 h at RT) ( ) and streptavidin-Alexafluor-488 (30 min; Invitrogen, Carlsbad, CA) were used. .. When biotinylated antibodies were used, a streptavidin-biotin blocking kit (Vector Laboratories, Burlingame, CA) was applied between each of the successive antibody incubations. .. The stained sections were mounted in antifading reagent (Slow Fade Gold; Invitrogen, Carlsbad, CA).

    Article Title: Loss of α2-6 sialylation promotes the transformation of synovial fibroblasts into a pro-inflammatory phenotype in Rheumatoid Arthritis
    Article Snippet: .. Endogenous biotin was blocked with the Streptavidin/Biotin blocking kit according to manufacturer’s instructions (Vector Laboratories, SP-2002). .. Carbo-Free Blocking Solution (Vector laboratories, SP-5040) was used as blocking solution.

    Article Title: Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro Confers Increased β1,6-N-Glycosylation and Increased Susceptibility to Galectin-3 Binding
    Article Snippet: In order to remove the paraffin and to rehydrate the sections, they were first rinsed with xylene (once for 5 min) and isopropanol (twice for 5 min), followed by ethanol (96% and 70%—each for 5 min) and finally millipore water (5 min). .. Prior to incubation with plant lectins, sections were blocked for 45 minutes in 1x carbohydrate-free blocking solution (Vector laboratories) and streptavidin blocking solution (Streptavidin/Biotin Blocking Kit, Vector laboratories) followed by addition of an appropriate amount of biotin solution for 15 minutes, in order to prevent non-specific staining. .. Sections were then incubated overnight at 4°C with 2 μg/mL biotin-conjugated PHA-L or biotin-conjugated ConA or streptavidin-peroxidase without prior addition of a plant lectin in order to visualize binding of biotinylated Gal-3 to native RPE cells in situ.

    Article Title: Antigen Display, T-Cell Activation, and Immune Evasion during Acute and Chronic Ehrlichiosis
    Article Snippet: The saponin-permeabilized cells were incubated with biotinylated Ec18.1 for 1 h at RT, followed by incubation with streptavidin-Alexafluor-488 (Invitrogen) for 30 min. .. Since all of the antibodies used in this staining procedure were biotinylated, a streptavidin-biotin blocking kit (Vector Laboratories, Burlingame, CA) was used between successive antibody incubations. .. For the detection of bacteria within peritoneal macrophages, the cells were stained with fluorescein isothiocyanate-F4/80 (Serotec) and biotinylated Ec18.1.

    Article Title: Loss of Mammal-specific Tectorial Membrane Component Carcinoembryonic Antigen Cell Adhesion Molecule 16 (CEACAM16) Leads to Hearing Impairment at Low and High Frequencies *
    Article Snippet: .. Antigen retrieval was achieved by heating to ∼95 °C of the tissue sections in Target Retrieval Solution, pH 9 (DakoCytomation), for 30 min followed by cooling to room temperature for 20 min. After blockage of endogenous peroxidase and biotin by incubation with 3% H2 O2 , 10% methanol in PBS for 5 min at room temperature and by using the streptavidin/biotin blocking kit (Vector Laboratories; Linearis), respectively, sections were reacted with the biotinylated anti-CEACAM16 antibody 9D5 (4 μg/ml) for 2 h at room temperature or overnight at 4 °C followed by a 1-h incubation with horseradish peroxidase-coupled streptavidin (Sigma) and stained by incubation with 3-amino-9-ethylcarbazole. .. For immunofluorescence staining, the cochlea and the vestibular organ were fixed for 2 h in 2% freshly prepared formaldehyde, 125 m m sucrose in 100 m m PBS, pH 7.4, at room temperature followed by overnight incubation with 25% sucrose with 1 m m protease inhibitor Pefabloc SC (Roche Diagnostics) in PBS.

    Article Title: Quantifying the Biomechanics of Conception: L-Selectin-Mediated Blastocyst Implantation Mechanics with Engineered “Trophospheres”
    Article Snippet: Flow cytometry primary antibodies were anti-human L-selectin (CD62L) monoclonal (Clone #4G8) antibody BBA24 (R & D Systems, Minneapolis, MN) or isotype-matched control (clone MOPC-1; BD Biosciences, San Jose, CA), and the secondary antibody was Alexa Fluor 488 goat anti-mouse IgG1 (Invitrogen, Carlsbad, CA). .. Immunoreagents for immunohistochemistry were mouse IgG1 (clone MOPC-21; BD Pharmingen), L-selectin antibody (Clone #4G8; R & D Systems), biotinylated goat anti-mouse IgG (H+L) (Vector Laboratories, Burlingame, CA), streptavidin/biotin blocking kit, and streptavidin-Texas Red (Vector Laboratories). .. The MECA-79 rat IgM antibody (BD Biosciences) and 3′-sLeX tetrasaccharide (Sigma) were used in flow detachment studies.

    Article Title: Osteoarticular tissue infection and development of skeletal pathology in murine brucellosis
    Article Snippet: Nonspecific binding was blocked for 1 hour at room temperature using 2% BSA (Sigma), 2% normal non-immune donkey serum (Vector Labs), and 0.05% Tween-20 in 1×PBS. .. Sections were rinsed briefly with 1×PBS and additionally blocked using streptavidin-biotin blocking kit (#SP-2002, Vector Labs). .. Finally, slides were incubated overnight at 4°C with biotinylated antibodies to Brucella (1.0 μg/ml; #TC-7011, Tetracore Inc.).

    Article Title: Regulation of Expression of Microvillus Membrane Proteins by Estrogen in Baboon Fetal Ovarian Oocytes 1
    Article Snippet: Briefly, for detection with DAB, sections were treated with VectaStain Elite Kit (Vector Laboratories Inc., Burlingame, CA), rinsed, stained with DAB-imidazole-H2 O2 , counterstained with Harris hematoxylin, mounted in Cytoseal-XYL (Richard Allan Scientific, Kalamazoo, MI), and examined by light microscopy [ ]. .. For immunofluorescence analysis, before blocking with NHS or NGS, sections were incubated with Image-iT signal enhancer (Molecular Probes) and blocked for endogenous biotin with the Streptavidin/Biotin Blocking kit (Vector Laboratories Inc., Center Valley, PA). .. After treatment with streptavidin-conjugated Alexa Fluor 488 diluted 1:500 with 5% NGS or NHS in PBS, sections were incubated with filtered 1% Sudan Black (Sigma-Aldrich) in 70% methanol to quench autofluorescence, rinsed, and treated with propidium iodide (1.0 μg/ml PBS) to stain nuclei red.

    Incubation:

    Article Title: Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro Confers Increased β1,6-N-Glycosylation and Increased Susceptibility to Galectin-3 Binding
    Article Snippet: In order to remove the paraffin and to rehydrate the sections, they were first rinsed with xylene (once for 5 min) and isopropanol (twice for 5 min), followed by ethanol (96% and 70%—each for 5 min) and finally millipore water (5 min). .. Prior to incubation with plant lectins, sections were blocked for 45 minutes in 1x carbohydrate-free blocking solution (Vector laboratories) and streptavidin blocking solution (Streptavidin/Biotin Blocking Kit, Vector laboratories) followed by addition of an appropriate amount of biotin solution for 15 minutes, in order to prevent non-specific staining. .. Sections were then incubated overnight at 4°C with 2 μg/mL biotin-conjugated PHA-L or biotin-conjugated ConA or streptavidin-peroxidase without prior addition of a plant lectin in order to visualize binding of biotinylated Gal-3 to native RPE cells in situ.

    Article Title: Loss of Mammal-specific Tectorial Membrane Component Carcinoembryonic Antigen Cell Adhesion Molecule 16 (CEACAM16) Leads to Hearing Impairment at Low and High Frequencies *
    Article Snippet: .. Antigen retrieval was achieved by heating to ∼95 °C of the tissue sections in Target Retrieval Solution, pH 9 (DakoCytomation), for 30 min followed by cooling to room temperature for 20 min. After blockage of endogenous peroxidase and biotin by incubation with 3% H2 O2 , 10% methanol in PBS for 5 min at room temperature and by using the streptavidin/biotin blocking kit (Vector Laboratories; Linearis), respectively, sections were reacted with the biotinylated anti-CEACAM16 antibody 9D5 (4 μg/ml) for 2 h at room temperature or overnight at 4 °C followed by a 1-h incubation with horseradish peroxidase-coupled streptavidin (Sigma) and stained by incubation with 3-amino-9-ethylcarbazole. .. For immunofluorescence staining, the cochlea and the vestibular organ were fixed for 2 h in 2% freshly prepared formaldehyde, 125 m m sucrose in 100 m m PBS, pH 7.4, at room temperature followed by overnight incubation with 25% sucrose with 1 m m protease inhibitor Pefabloc SC (Roche Diagnostics) in PBS.

    Article Title: Regulation of Expression of Microvillus Membrane Proteins by Estrogen in Baboon Fetal Ovarian Oocytes 1
    Article Snippet: Briefly, for detection with DAB, sections were treated with VectaStain Elite Kit (Vector Laboratories Inc., Burlingame, CA), rinsed, stained with DAB-imidazole-H2 O2 , counterstained with Harris hematoxylin, mounted in Cytoseal-XYL (Richard Allan Scientific, Kalamazoo, MI), and examined by light microscopy [ ]. .. For immunofluorescence analysis, before blocking with NHS or NGS, sections were incubated with Image-iT signal enhancer (Molecular Probes) and blocked for endogenous biotin with the Streptavidin/Biotin Blocking kit (Vector Laboratories Inc., Center Valley, PA). .. After treatment with streptavidin-conjugated Alexa Fluor 488 diluted 1:500 with 5% NGS or NHS in PBS, sections were incubated with filtered 1% Sudan Black (Sigma-Aldrich) in 70% methanol to quench autofluorescence, rinsed, and treated with propidium iodide (1.0 μg/ml PBS) to stain nuclei red.

    Staining:

    Article Title: Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro Confers Increased β1,6-N-Glycosylation and Increased Susceptibility to Galectin-3 Binding
    Article Snippet: In order to remove the paraffin and to rehydrate the sections, they were first rinsed with xylene (once for 5 min) and isopropanol (twice for 5 min), followed by ethanol (96% and 70%—each for 5 min) and finally millipore water (5 min). .. Prior to incubation with plant lectins, sections were blocked for 45 minutes in 1x carbohydrate-free blocking solution (Vector laboratories) and streptavidin blocking solution (Streptavidin/Biotin Blocking Kit, Vector laboratories) followed by addition of an appropriate amount of biotin solution for 15 minutes, in order to prevent non-specific staining. .. Sections were then incubated overnight at 4°C with 2 μg/mL biotin-conjugated PHA-L or biotin-conjugated ConA or streptavidin-peroxidase without prior addition of a plant lectin in order to visualize binding of biotinylated Gal-3 to native RPE cells in situ.

    Article Title: Antigen Display, T-Cell Activation, and Immune Evasion during Acute and Chronic Ehrlichiosis
    Article Snippet: The saponin-permeabilized cells were incubated with biotinylated Ec18.1 for 1 h at RT, followed by incubation with streptavidin-Alexafluor-488 (Invitrogen) for 30 min. .. Since all of the antibodies used in this staining procedure were biotinylated, a streptavidin-biotin blocking kit (Vector Laboratories, Burlingame, CA) was used between successive antibody incubations. .. For the detection of bacteria within peritoneal macrophages, the cells were stained with fluorescein isothiocyanate-F4/80 (Serotec) and biotinylated Ec18.1.

    Article Title: Loss of Mammal-specific Tectorial Membrane Component Carcinoembryonic Antigen Cell Adhesion Molecule 16 (CEACAM16) Leads to Hearing Impairment at Low and High Frequencies *
    Article Snippet: .. Antigen retrieval was achieved by heating to ∼95 °C of the tissue sections in Target Retrieval Solution, pH 9 (DakoCytomation), for 30 min followed by cooling to room temperature for 20 min. After blockage of endogenous peroxidase and biotin by incubation with 3% H2 O2 , 10% methanol in PBS for 5 min at room temperature and by using the streptavidin/biotin blocking kit (Vector Laboratories; Linearis), respectively, sections were reacted with the biotinylated anti-CEACAM16 antibody 9D5 (4 μg/ml) for 2 h at room temperature or overnight at 4 °C followed by a 1-h incubation with horseradish peroxidase-coupled streptavidin (Sigma) and stained by incubation with 3-amino-9-ethylcarbazole. .. For immunofluorescence staining, the cochlea and the vestibular organ were fixed for 2 h in 2% freshly prepared formaldehyde, 125 m m sucrose in 100 m m PBS, pH 7.4, at room temperature followed by overnight incubation with 25% sucrose with 1 m m protease inhibitor Pefabloc SC (Roche Diagnostics) in PBS.

    Immunohistochemistry:

    Article Title: Quantifying the Biomechanics of Conception: L-Selectin-Mediated Blastocyst Implantation Mechanics with Engineered “Trophospheres”
    Article Snippet: Flow cytometry primary antibodies were anti-human L-selectin (CD62L) monoclonal (Clone #4G8) antibody BBA24 (R & D Systems, Minneapolis, MN) or isotype-matched control (clone MOPC-1; BD Biosciences, San Jose, CA), and the secondary antibody was Alexa Fluor 488 goat anti-mouse IgG1 (Invitrogen, Carlsbad, CA). .. Immunoreagents for immunohistochemistry were mouse IgG1 (clone MOPC-21; BD Pharmingen), L-selectin antibody (Clone #4G8; R & D Systems), biotinylated goat anti-mouse IgG (H+L) (Vector Laboratories, Burlingame, CA), streptavidin/biotin blocking kit, and streptavidin-Texas Red (Vector Laboratories). .. The MECA-79 rat IgM antibody (BD Biosciences) and 3′-sLeX tetrasaccharide (Sigma) were used in flow detachment studies.

    Immunofluorescence:

    Article Title: Regulation of Expression of Microvillus Membrane Proteins by Estrogen in Baboon Fetal Ovarian Oocytes 1
    Article Snippet: Briefly, for detection with DAB, sections were treated with VectaStain Elite Kit (Vector Laboratories Inc., Burlingame, CA), rinsed, stained with DAB-imidazole-H2 O2 , counterstained with Harris hematoxylin, mounted in Cytoseal-XYL (Richard Allan Scientific, Kalamazoo, MI), and examined by light microscopy [ ]. .. For immunofluorescence analysis, before blocking with NHS or NGS, sections were incubated with Image-iT signal enhancer (Molecular Probes) and blocked for endogenous biotin with the Streptavidin/Biotin Blocking kit (Vector Laboratories Inc., Center Valley, PA). .. After treatment with streptavidin-conjugated Alexa Fluor 488 diluted 1:500 with 5% NGS or NHS in PBS, sections were incubated with filtered 1% Sudan Black (Sigma-Aldrich) in 70% methanol to quench autofluorescence, rinsed, and treated with propidium iodide (1.0 μg/ml PBS) to stain nuclei red.

    Next-Generation Sequencing:

    Article Title: Regulation of Expression of Microvillus Membrane Proteins by Estrogen in Baboon Fetal Ovarian Oocytes 1
    Article Snippet: Briefly, for detection with DAB, sections were treated with VectaStain Elite Kit (Vector Laboratories Inc., Burlingame, CA), rinsed, stained with DAB-imidazole-H2 O2 , counterstained with Harris hematoxylin, mounted in Cytoseal-XYL (Richard Allan Scientific, Kalamazoo, MI), and examined by light microscopy [ ]. .. For immunofluorescence analysis, before blocking with NHS or NGS, sections were incubated with Image-iT signal enhancer (Molecular Probes) and blocked for endogenous biotin with the Streptavidin/Biotin Blocking kit (Vector Laboratories Inc., Center Valley, PA). .. After treatment with streptavidin-conjugated Alexa Fluor 488 diluted 1:500 with 5% NGS or NHS in PBS, sections were incubated with filtered 1% Sudan Black (Sigma-Aldrich) in 70% methanol to quench autofluorescence, rinsed, and treated with propidium iodide (1.0 μg/ml PBS) to stain nuclei red.

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    Vector Laboratories streptavidin biotin blocking kit
    Expression of CEACAM16 in the cochlea and vestibular organ. Expression of CEACAM16 was analyzed in the cochleae and vestibular organ of Ceacam16 +/+ ( A–C , I , and J ) and Ceacam16 −/− mice ( E–G ) by immunohistology using the biotinylated monoclonal anti-human CEACAM16 antibody 9D5, which cross-reacts with murine CEACAM16. CEACAM16 ( brown stain ) was observed in the acellular TM ( A , B , I , and J ) and in the extracellular matrix covering the macula cells of the saccule ( C ) in wild-type but not in Ceacam16 −/− mice. Furthermore, staining for CEACAM16 was observed in interdental cells in the helicotrema region of P15 wild-type cochleae ( I ) and also in adult Ceacam16 +/− mice ( J ). No staining was seen when only horseradish peroxidase-coupled <t>streptavidin</t> was added ( K ). Comparable staining for α-tectorin was found in the TM in both Ceacam16 +/+ ( D ) and Ceacam16 −/− mice ( H ). The age of the mice (days post partum) is indicated at the lower left corners . Note that the TM of Ceacam16 −/− mice is significantly more often in contact with the OHCs ( E , F , and H ), whereas it is either detached or more retracted in wild-type mice ( B , D , I , and J ). No difference was observed when different regions of the cochlea (base, medal region, or helicotrema-proximal) were analyzed. Five wild-type and four Ceacam16 −/− mice, representing eight and seven cochleae, respectively, were evaluated; p = 0.0003, Fisher's exact test (L). B , basilar membrane; GER , greater epithelial ridge; ID , interdental cells; IS , inner sulcus; M , macula; R , Reissner's membrane; SV , stria vascularis; SL , spiral limbus; TM , tectorial membrane.
    Streptavidin Biotin Blocking Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin biotin blocking kit/product/Vector Laboratories
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin biotin blocking kit - by Bioz Stars, 2021-03
    94/100 stars
      Buy from Supplier

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    Expression of CEACAM16 in the cochlea and vestibular organ. Expression of CEACAM16 was analyzed in the cochleae and vestibular organ of Ceacam16 +/+ ( A–C , I , and J ) and Ceacam16 −/− mice ( E–G ) by immunohistology using the biotinylated monoclonal anti-human CEACAM16 antibody 9D5, which cross-reacts with murine CEACAM16. CEACAM16 ( brown stain ) was observed in the acellular TM ( A , B , I , and J ) and in the extracellular matrix covering the macula cells of the saccule ( C ) in wild-type but not in Ceacam16 −/− mice. Furthermore, staining for CEACAM16 was observed in interdental cells in the helicotrema region of P15 wild-type cochleae ( I ) and also in adult Ceacam16 +/− mice ( J ). No staining was seen when only horseradish peroxidase-coupled streptavidin was added ( K ). Comparable staining for α-tectorin was found in the TM in both Ceacam16 +/+ ( D ) and Ceacam16 −/− mice ( H ). The age of the mice (days post partum) is indicated at the lower left corners . Note that the TM of Ceacam16 −/− mice is significantly more often in contact with the OHCs ( E , F , and H ), whereas it is either detached or more retracted in wild-type mice ( B , D , I , and J ). No difference was observed when different regions of the cochlea (base, medal region, or helicotrema-proximal) were analyzed. Five wild-type and four Ceacam16 −/− mice, representing eight and seven cochleae, respectively, were evaluated; p = 0.0003, Fisher's exact test (L). B , basilar membrane; GER , greater epithelial ridge; ID , interdental cells; IS , inner sulcus; M , macula; R , Reissner's membrane; SV , stria vascularis; SL , spiral limbus; TM , tectorial membrane.

    Journal: The Journal of Biological Chemistry

    Article Title: Loss of Mammal-specific Tectorial Membrane Component Carcinoembryonic Antigen Cell Adhesion Molecule 16 (CEACAM16) Leads to Hearing Impairment at Low and High Frequencies *

    doi: 10.1074/jbc.M111.320481

    Figure Lengend Snippet: Expression of CEACAM16 in the cochlea and vestibular organ. Expression of CEACAM16 was analyzed in the cochleae and vestibular organ of Ceacam16 +/+ ( A–C , I , and J ) and Ceacam16 −/− mice ( E–G ) by immunohistology using the biotinylated monoclonal anti-human CEACAM16 antibody 9D5, which cross-reacts with murine CEACAM16. CEACAM16 ( brown stain ) was observed in the acellular TM ( A , B , I , and J ) and in the extracellular matrix covering the macula cells of the saccule ( C ) in wild-type but not in Ceacam16 −/− mice. Furthermore, staining for CEACAM16 was observed in interdental cells in the helicotrema region of P15 wild-type cochleae ( I ) and also in adult Ceacam16 +/− mice ( J ). No staining was seen when only horseradish peroxidase-coupled streptavidin was added ( K ). Comparable staining for α-tectorin was found in the TM in both Ceacam16 +/+ ( D ) and Ceacam16 −/− mice ( H ). The age of the mice (days post partum) is indicated at the lower left corners . Note that the TM of Ceacam16 −/− mice is significantly more often in contact with the OHCs ( E , F , and H ), whereas it is either detached or more retracted in wild-type mice ( B , D , I , and J ). No difference was observed when different regions of the cochlea (base, medal region, or helicotrema-proximal) were analyzed. Five wild-type and four Ceacam16 −/− mice, representing eight and seven cochleae, respectively, were evaluated; p = 0.0003, Fisher's exact test (L). B , basilar membrane; GER , greater epithelial ridge; ID , interdental cells; IS , inner sulcus; M , macula; R , Reissner's membrane; SV , stria vascularis; SL , spiral limbus; TM , tectorial membrane.

    Article Snippet: Antigen retrieval was achieved by heating to ∼95 °C of the tissue sections in Target Retrieval Solution, pH 9 (DakoCytomation), for 30 min followed by cooling to room temperature for 20 min. After blockage of endogenous peroxidase and biotin by incubation with 3% H2 O2 , 10% methanol in PBS for 5 min at room temperature and by using the streptavidin/biotin blocking kit (Vector Laboratories; Linearis), respectively, sections were reacted with the biotinylated anti-CEACAM16 antibody 9D5 (4 μg/ml) for 2 h at room temperature or overnight at 4 °C followed by a 1-h incubation with horseradish peroxidase-coupled streptavidin (Sigma) and stained by incubation with 3-amino-9-ethylcarbazole.

    Techniques: Expressing, Mouse Assay, Staining

    Trophosphere imaging. (A) Scanning electron micrograph of a trophosphere. (B) L-selectin expression was analyzed by confocal microscopy with trophospheres stained with Dreg-56, biotinylated secondary antibody, and Texas Red-conjugated streptavidin (200×). (C, D) Representative light micrographs (100×) of trophospheres used for dimensional analysis. Diameter, circularity, and roundness measurements are (C, 238 μm, 0.77, 0.92 ) and (D, 299 μm, 0.84, 0.85 )

    Journal: Tissue Engineering. Part A

    Article Title: Quantifying the Biomechanics of Conception: L-Selectin-Mediated Blastocyst Implantation Mechanics with Engineered “Trophospheres”

    doi: 10.1089/ten.tea.2013.0067

    Figure Lengend Snippet: Trophosphere imaging. (A) Scanning electron micrograph of a trophosphere. (B) L-selectin expression was analyzed by confocal microscopy with trophospheres stained with Dreg-56, biotinylated secondary antibody, and Texas Red-conjugated streptavidin (200×). (C, D) Representative light micrographs (100×) of trophospheres used for dimensional analysis. Diameter, circularity, and roundness measurements are (C, 238 μm, 0.77, 0.92 ) and (D, 299 μm, 0.84, 0.85 )

    Article Snippet: Immunoreagents for immunohistochemistry were mouse IgG1 (clone MOPC-21; BD Pharmingen), L-selectin antibody (Clone #4G8; R & D Systems), biotinylated goat anti-mouse IgG (H+L) (Vector Laboratories, Burlingame, CA), streptavidin/biotin blocking kit, and streptavidin-Texas Red (Vector Laboratories).

    Techniques: Imaging, Expressing, Confocal Microscopy, Staining

    DCs display antigen and harbor E. muris . DCs were purified from pooled day 5-infected spleens and LNs ( n = 3), enriched by magnetic bead positive selection, and then purified by flow-cytometric cell sorting (95 to 99% purity was obtained). (a) Representative dot plots demonstrating the homogeneity of each of the purified DC populations. (b) OMP-19 107-122 antigen presentation was measured in cultures containing the DCs, purified as described for panel a, from mock-infected mice (mock) or from spleen and LNs from mice on day 5 postinfection. The data are representative of two independent experiments of similar design; the average frequencies of IFN-γ-producing T cells were 43.7 and 13.6% for spleen and LNs, respectively. (c) DCs from mice on day 5 postinfection were purified by flow-cytometric cell sorting as shown in panel a and were stained with antibodies that recognize E. muris (Ec18.1) and CD11c. The staining for E. muris was performed using biotinylated Ec18.1. The cells first were stained using streptavidin-conjugated Alexafluor-594 (pseudocolored cyan in the figure), permeabilized with 0.2% saponin, blocked, and stained again with streptavidin-conjugated Alexafluor-488 (shown in green). Thus, the cyan-colored bacteria are surface-associated bacteria, and the green bacteria represent all of the bacteria associated with the host cell. The merged image, which also includes CD11c staining (in red), is shown in the panels at the right. Nuclei were counterstained with 4′,6′-diamidino-2-phenylindole (blue in nuclei). The top and bottom rows show two representative fields of cells.

    Journal: Infection and Immunity

    Article Title: Antigen Display, T-Cell Activation, and Immune Evasion during Acute and Chronic Ehrlichiosis

    doi: 10.1128/IAI.01433-08

    Figure Lengend Snippet: DCs display antigen and harbor E. muris . DCs were purified from pooled day 5-infected spleens and LNs ( n = 3), enriched by magnetic bead positive selection, and then purified by flow-cytometric cell sorting (95 to 99% purity was obtained). (a) Representative dot plots demonstrating the homogeneity of each of the purified DC populations. (b) OMP-19 107-122 antigen presentation was measured in cultures containing the DCs, purified as described for panel a, from mock-infected mice (mock) or from spleen and LNs from mice on day 5 postinfection. The data are representative of two independent experiments of similar design; the average frequencies of IFN-γ-producing T cells were 43.7 and 13.6% for spleen and LNs, respectively. (c) DCs from mice on day 5 postinfection were purified by flow-cytometric cell sorting as shown in panel a and were stained with antibodies that recognize E. muris (Ec18.1) and CD11c. The staining for E. muris was performed using biotinylated Ec18.1. The cells first were stained using streptavidin-conjugated Alexafluor-594 (pseudocolored cyan in the figure), permeabilized with 0.2% saponin, blocked, and stained again with streptavidin-conjugated Alexafluor-488 (shown in green). Thus, the cyan-colored bacteria are surface-associated bacteria, and the green bacteria represent all of the bacteria associated with the host cell. The merged image, which also includes CD11c staining (in red), is shown in the panels at the right. Nuclei were counterstained with 4′,6′-diamidino-2-phenylindole (blue in nuclei). The top and bottom rows show two representative fields of cells.

    Article Snippet: When biotinylated antibodies were used, a streptavidin-biotin blocking kit (Vector Laboratories, Burlingame, CA) was applied between each of the successive antibody incubations.

    Techniques: Purification, Infection, Selection, Flow Cytometry, FACS, Mouse Assay, Staining

    Representative photomicrographs of the immunocytochemical expression of α-actinin in oocytes and pregranulosa cells in the fetal ovary of untreated baboons on Day 100 of gestation ( A ) and in oocytes of primordial follicles on Day 165 ( B – D ) of gestation in animals untreated ( B ) or treated with CGS 20267 (115 μg/kg body weight per day; C and inset of C ) or CGS 20267 and estradiol benzoate (115 μg/kg body weight per day each; D and inset of D ) administered s.c. to the mother beginning on Day 100 (term = Day 184). E ) Expression of α-actinin in blood vessels in fetal ovary at Days 100 (panel 1) and 165 (panel 2) of gestation in untreated baboons, on Day 165 in animals treated with CGS 20267 (panel 3), and in adult ovary (panel 4). Sections were incubated with antibody to α-actinin, stained with streptavidin conjugated with AlexaFluor 488 (green), and treated with propidium iodide to stain nuclei (red). F ) Section of late-gestation untreated fetal ovary incubated without primary antibody. Original magnifications ×400 ( A – D ); ×1000 ( B – D , insets); and ×400 ( E , F ). gc, Granulosa cell; n, oocyte nucleus. Bars = 25 μm ( A – E ) and 10 μm ( B – D , insets).

    Journal: Biology of Reproduction

    Article Title: Regulation of Expression of Microvillus Membrane Proteins by Estrogen in Baboon Fetal Ovarian Oocytes 1

    doi: 10.1095/biolreprod.108.067900

    Figure Lengend Snippet: Representative photomicrographs of the immunocytochemical expression of α-actinin in oocytes and pregranulosa cells in the fetal ovary of untreated baboons on Day 100 of gestation ( A ) and in oocytes of primordial follicles on Day 165 ( B – D ) of gestation in animals untreated ( B ) or treated with CGS 20267 (115 μg/kg body weight per day; C and inset of C ) or CGS 20267 and estradiol benzoate (115 μg/kg body weight per day each; D and inset of D ) administered s.c. to the mother beginning on Day 100 (term = Day 184). E ) Expression of α-actinin in blood vessels in fetal ovary at Days 100 (panel 1) and 165 (panel 2) of gestation in untreated baboons, on Day 165 in animals treated with CGS 20267 (panel 3), and in adult ovary (panel 4). Sections were incubated with antibody to α-actinin, stained with streptavidin conjugated with AlexaFluor 488 (green), and treated with propidium iodide to stain nuclei (red). F ) Section of late-gestation untreated fetal ovary incubated without primary antibody. Original magnifications ×400 ( A – D ); ×1000 ( B – D , insets); and ×400 ( E , F ). gc, Granulosa cell; n, oocyte nucleus. Bars = 25 μm ( A – E ) and 10 μm ( B – D , insets).

    Article Snippet: For immunofluorescence analysis, before blocking with NHS or NGS, sections were incubated with Image-iT signal enhancer (Molecular Probes) and blocked for endogenous biotin with the Streptavidin/Biotin Blocking kit (Vector Laboratories Inc., Center Valley, PA).

    Techniques: Expressing, Incubation, Staining