rabbit anti goat alexa 488 antibody  (Thermo Fisher)


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    Name:
    Rabbit anti Goat IgG H L Superclonal Secondary Antibody
    Description:
    Rabbit anti Goat IgG H L Secondary Antibody for Western Blot IF ICC ELISA
    Catalog Number:
    A27011
    Price:
    None
    Category:
    Antibodies Secondary Detection Reagents
    Applications:
    Build Your Own Immunoassay|Cell Analysis|Cellular Imaging|ELISA|Flow Cytometry Antibodies & Secondary Detection|Fluorescence Microscopy|Immunocytochemistry (ICC)|Microscopy|Protein Assays and Analysis|Protein Biology|Ready-To-Use Immunoassay|Secondary Detection|Flow Cytometry
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    Structured Review

    Thermo Fisher rabbit anti goat alexa 488 antibody
    Establishment of the optimal printing and storage conditions for biotinylated antigen arrays in streptavidin-coated 96-well plates. (A) Schematic representation of a 96 well plate with a 5 × 5 antigen array. (B–L) A purified biotinylated antibody (OX68-bio) was spotted in streptavidin-coated 96 well plates. (B and C) Detection with either a non-fluorescent alkaline phosphatase-conjugated secondary antibody followed by a precipitating colourimetric NBT/BCIP substrate (B); and, with a fluorescent Alexa 568-conjugated secondary antibody (C). (D) Different concentrations of OX68-bio, 1 = 0.5; 2 = 0.25; 3 = 0.1; 4 = 0.05 μg/μl were arranged vertically, and in quadruplicate horizontally. (E) Different amounts of Tween 20 added prior to printing, 1 = 10%; 2 = 1%; 3 = 0.1%; 4 = None. (F) Different amounts of glycerol added prior to printing, 1 = 10%; 2 = 1%; 3 = 0.1%; 4 = 1% Tween 20. (G) Incubation of different concentrations of spotted antibody (as in (D)) for 2 h at room temperature with a 4% formalin solution. Storage of a printed plate for 2 days at room temperature (H), and at −20 °C (I), prior to antigen detection. Examples of: a 25-spot array using 0.4 mm pins, 0.66 mm pitch (J); a 53-spot array using 0.2 mm pins, 0.45 mm pitch (K), and a 101-spot array using 0.2 mm pins, 0.32 mm pitch. In panels D–L printed antibody was detected with an <t>Alexa-488</t> conjugated secondary antibody.
    Rabbit anti Goat IgG H L Secondary Antibody for Western Blot IF ICC ELISA
    https://www.bioz.com/result/rabbit anti goat alexa 488 antibody/product/Thermo Fisher
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti goat alexa 488 antibody - by Bioz Stars, 2021-04
    94/100 stars

    Images

    1) Product Images from "Development of an antigen microarray for high throughput monoclonal antibody selection"

    Article Title: Development of an antigen microarray for high throughput monoclonal antibody selection

    Journal: Biochemical and Biophysical Research Communications

    doi: 10.1016/j.bbrc.2013.12.033

    Establishment of the optimal printing and storage conditions for biotinylated antigen arrays in streptavidin-coated 96-well plates. (A) Schematic representation of a 96 well plate with a 5 × 5 antigen array. (B–L) A purified biotinylated antibody (OX68-bio) was spotted in streptavidin-coated 96 well plates. (B and C) Detection with either a non-fluorescent alkaline phosphatase-conjugated secondary antibody followed by a precipitating colourimetric NBT/BCIP substrate (B); and, with a fluorescent Alexa 568-conjugated secondary antibody (C). (D) Different concentrations of OX68-bio, 1 = 0.5; 2 = 0.25; 3 = 0.1; 4 = 0.05 μg/μl were arranged vertically, and in quadruplicate horizontally. (E) Different amounts of Tween 20 added prior to printing, 1 = 10%; 2 = 1%; 3 = 0.1%; 4 = None. (F) Different amounts of glycerol added prior to printing, 1 = 10%; 2 = 1%; 3 = 0.1%; 4 = 1% Tween 20. (G) Incubation of different concentrations of spotted antibody (as in (D)) for 2 h at room temperature with a 4% formalin solution. Storage of a printed plate for 2 days at room temperature (H), and at −20 °C (I), prior to antigen detection. Examples of: a 25-spot array using 0.4 mm pins, 0.66 mm pitch (J); a 53-spot array using 0.2 mm pins, 0.45 mm pitch (K), and a 101-spot array using 0.2 mm pins, 0.32 mm pitch. In panels D–L printed antibody was detected with an Alexa-488 conjugated secondary antibody.
    Figure Legend Snippet: Establishment of the optimal printing and storage conditions for biotinylated antigen arrays in streptavidin-coated 96-well plates. (A) Schematic representation of a 96 well plate with a 5 × 5 antigen array. (B–L) A purified biotinylated antibody (OX68-bio) was spotted in streptavidin-coated 96 well plates. (B and C) Detection with either a non-fluorescent alkaline phosphatase-conjugated secondary antibody followed by a precipitating colourimetric NBT/BCIP substrate (B); and, with a fluorescent Alexa 568-conjugated secondary antibody (C). (D) Different concentrations of OX68-bio, 1 = 0.5; 2 = 0.25; 3 = 0.1; 4 = 0.05 μg/μl were arranged vertically, and in quadruplicate horizontally. (E) Different amounts of Tween 20 added prior to printing, 1 = 10%; 2 = 1%; 3 = 0.1%; 4 = None. (F) Different amounts of glycerol added prior to printing, 1 = 10%; 2 = 1%; 3 = 0.1%; 4 = 1% Tween 20. (G) Incubation of different concentrations of spotted antibody (as in (D)) for 2 h at room temperature with a 4% formalin solution. Storage of a printed plate for 2 days at room temperature (H), and at −20 °C (I), prior to antigen detection. Examples of: a 25-spot array using 0.4 mm pins, 0.66 mm pitch (J); a 53-spot array using 0.2 mm pins, 0.45 mm pitch (K), and a 101-spot array using 0.2 mm pins, 0.32 mm pitch. In panels D–L printed antibody was detected with an Alexa-488 conjugated secondary antibody.

    Techniques Used: Purification, Incubation

    Related Articles

    Staining:

    Article Title: Protein Tyrosine Phosphatase Non-Receptor Type 2 Function in Dendritic Cells Is Crucial to Maintain Tissue Tolerance
    Article Snippet: For liver, lung, kidney, and spleen, erythrocyte lysis was performed using ammonium-chloride-potassium (ACK) buffer (150 mM NH4 Cl, 10 mM KHCO3 , 0.1 mM Na2EDTA). .. The samples were then stained with fluorescence cytometry antibodies. .. Flow Cytometry Cells were incubated with primary antibodies in PBS for 30 min at 4°C and washed with PBS.

    Article Title: Alteration of Lysosome Fusion and Low-grade Inflammation Mediated by Super-low-dose Endotoxin *
    Article Snippet: An increase in mean fluorescence intensity represents a decrease in acidification. .. BMDMs harvested from WT mice were seeded on coverslips and treated with or without super-low-dose LPS (50 pg/ml) for 1 or 24 h. Some samples were labeled with MitoTracker Red (200 n m ; Invitrogen) for 20 min. After fixation with 4% paraformaldehyde and permeabilization with 0.2% saponin, the cells were stained with goat anti-mouse Tollip antibody (Santa Cruz Biotechnology), followed by Alexa Fluor 488-labeled rabbit anti-goat IgG (Invitrogen). .. In some experiments, cells were also co-stained with Cy3-conjugated rabbit anti-mouse LAMP1 antibody (Abcam).

    Article Title: Development of new therapy for canine mammary cancer with recombinant measles virus
    Article Snippet: .. The cells were washed once and stained with 0.1 μg of Alexa-Fluor-488-conjugated rabbit anti-goat IgG antibody (Molecular Probes, Eugene, OR) in 100 μl of sample buffer on ice for 45 minutes. .. After the cells were washed, they were resuspended in PBS containing 7-aminoactinomycin D (Beckman Coulter Immunotech, Massielle, France).

    Fluorescence:

    Article Title: Protein Tyrosine Phosphatase Non-Receptor Type 2 Function in Dendritic Cells Is Crucial to Maintain Tissue Tolerance
    Article Snippet: For liver, lung, kidney, and spleen, erythrocyte lysis was performed using ammonium-chloride-potassium (ACK) buffer (150 mM NH4 Cl, 10 mM KHCO3 , 0.1 mM Na2EDTA). .. The samples were then stained with fluorescence cytometry antibodies. .. Flow Cytometry Cells were incubated with primary antibodies in PBS for 30 min at 4°C and washed with PBS.

    Cytometry:

    Article Title: Protein Tyrosine Phosphatase Non-Receptor Type 2 Function in Dendritic Cells Is Crucial to Maintain Tissue Tolerance
    Article Snippet: For liver, lung, kidney, and spleen, erythrocyte lysis was performed using ammonium-chloride-potassium (ACK) buffer (150 mM NH4 Cl, 10 mM KHCO3 , 0.1 mM Na2EDTA). .. The samples were then stained with fluorescence cytometry antibodies. .. Flow Cytometry Cells were incubated with primary antibodies in PBS for 30 min at 4°C and washed with PBS.

    Immunohistochemistry:

    Article Title: Unacylated ghrelin promotes adipogenesis in rodent bone marrow via ghrelin O-acyl transferase and GHS-R1a activity: evidence for target cell-induced acylation
    Article Snippet: GOAT-positive cells in the marrow cell smears were identified by double fluorescence IHC for GOAT and PPARγ. .. The above IHC protocol was followed except that the primary antibody solution contained both rabbit anti-GOAT and mouse anti-human PPARγ (1:100 dilution; Cat No 419300; Invitrogen Corporation, Camarillo, CA; ) and the secondary antibody solution contained both Cy3-linked sheep anti-rabbit and Alexa-fluor488-linked goat anti-mouse antibodies (1:500). .. Study 4: Visualisation of the subcellular distribution of GOAT in tibial bone marrow adipocytes The subcellular distribution of GOAT in marrow adipocytes was visualised by immunogold electron microscopy (EM).

    Incubation:

    Article Title: Development of an antigen microarray for high throughput monoclonal antibody selection
    Article Snippet: 2.4 Screening of protein microarrays Hybridoma supernatants were added to the antigen arrays and incubated overnight at 4 °C. .. After washing with PBT (PBS + 0.1% Tween), arrays were incubated for 2 hours with a goat anti-mouse Alexa 488 secondary antibody (Invitrogen), washed in PBT, followed by a rabbit anti-goat Alexa 488 antibody (Invitrogen). .. This second amplification step, although not absolutely required, increased signals to permit direct visual screening using an epifluorescence microscope.

    Article Title: Adapter Protein SH2-B? Undergoes Nucleocytoplasmic Shuttling: Implications for Nerve Growth Factor Induction of Neuronal Differentiation
    Article Snippet: .. Fixed cells were incubated with anti-SH2-Bβ antibody (Santa Cruz) (diluted 1:25) for 1 h, with rabbit anti-goat Alexa 488 antibody (Fab′) (Molecular Probe) (diluted 1:100) for 1 h, and then with 4′,6′-diamidino-2-phenylindole (DAPI) (Molecular Probe) (2 ng/ml) for 5 min. .. Active ERK was immunostained with active ERK antibody (diluted 1:250) (Promega) for 1 h, with goat anti-rabbit Alexa 555 antibody (Molecular Probe) (diluted 1:500) for 1 h, and with 2 ng of DAPI per ml for 5 min.

    Mouse Assay:

    Article Title: Alteration of Lysosome Fusion and Low-grade Inflammation Mediated by Super-low-dose Endotoxin *
    Article Snippet: An increase in mean fluorescence intensity represents a decrease in acidification. .. BMDMs harvested from WT mice were seeded on coverslips and treated with or without super-low-dose LPS (50 pg/ml) for 1 or 24 h. Some samples were labeled with MitoTracker Red (200 n m ; Invitrogen) for 20 min. After fixation with 4% paraformaldehyde and permeabilization with 0.2% saponin, the cells were stained with goat anti-mouse Tollip antibody (Santa Cruz Biotechnology), followed by Alexa Fluor 488-labeled rabbit anti-goat IgG (Invitrogen). .. In some experiments, cells were also co-stained with Cy3-conjugated rabbit anti-mouse LAMP1 antibody (Abcam).

    Labeling:

    Article Title: Alteration of Lysosome Fusion and Low-grade Inflammation Mediated by Super-low-dose Endotoxin *
    Article Snippet: An increase in mean fluorescence intensity represents a decrease in acidification. .. BMDMs harvested from WT mice were seeded on coverslips and treated with or without super-low-dose LPS (50 pg/ml) for 1 or 24 h. Some samples were labeled with MitoTracker Red (200 n m ; Invitrogen) for 20 min. After fixation with 4% paraformaldehyde and permeabilization with 0.2% saponin, the cells were stained with goat anti-mouse Tollip antibody (Santa Cruz Biotechnology), followed by Alexa Fluor 488-labeled rabbit anti-goat IgG (Invitrogen). .. In some experiments, cells were also co-stained with Cy3-conjugated rabbit anti-mouse LAMP1 antibody (Abcam).

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  • 97
    Thermo Fisher goat anti rabbit igg alexa fluor 488
    Expression of integrin α 6 β 1 and stiffness of OPCs at different pH. ( a ) Expression level of integrin α 6 β 1 at different extracellular pH, evaluated by OPC immunostaining against α 6 β 1 (with <t>Alexa</t> <t>Fluor-488</t> fluorochrome) and analysis of cell fluorescence using flow cytometry (BD LSR Fortessa). Data are geometric mean fluorescence intensities averaged over three experiments, each conducted in triplicate, and presented relative to value obtained for pH 7.0. No statistical difference was observed between any pH conditions. ( b ) Cell stiffness at pH 6.0 and 7.0, evaluated using AFM-enabled nanoindentation. Data are mean of Young's elastic modulus measured for 15 cells per condition. No statistical difference was observed between pH 6.0 and 7.0. Error bars are SEM. Colors correspond to cell media pH.
    Goat Anti Rabbit Igg Alexa Fluor 488, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit igg alexa fluor 488/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg alexa fluor 488 - by Bioz Stars, 2021-04
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    99
    Thermo Fisher alexa 488 conjugated goat anti rabbit antibody
    Photolyase activation prevents UVB-induced DNA damage and restores cell viability HaCaT keratinocytes (n = 2 × 10 5 cell) were treated with 20 or 40 mJ/cm 2 UVB, then cells were harvested. CPD formation from (a) genomic and (b) mitochondrial DNA was measured by CPD ELISA (n = 3) after UVB exposure, as indicated. (c) Cell viability was assessed by Annexin <t>V-Alexa-488</t> and Propidium iodide labeling (n = 5) 24 after UVB exposure. (d) Cells were harvested 24 h post-irradiation and a clonogenic assay was performed (5000 cells/dish, n=4). (e) Cell cycle progression was assessed by Propidium Iodide incorporation (n = 4) 24 after UVB exposure. - represents where photolyase is inactive, + represents where photolyase is active *; **, and *** indicate statistically significant differences at p
    Alexa 488 Conjugated Goat Anti Rabbit Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alexa 488 conjugated goat anti rabbit antibody/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alexa 488 conjugated goat anti rabbit antibody - by Bioz Stars, 2021-04
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    97
    Thermo Fisher goat anti rabbit alexa fluor 488 antibodies
    Tick persistence depends on rel Bbu . (A) Immunofluorescence microscopy of ticks allowed to feed on mice infected with wild-type (WT), rel Bbu mutant ( rel Bbu - ) or rel Bbu - pBS- rel Bbu strains. Ticks were dissected and fixed on slides one week after naïve larvae had fed to repletion (fed larvae) or after larvae had molted to nymphs (unfed nymph) or one week after nymphs had fed to repletion on uninfected mice (fed nymph). Samples were processed for IF microscopy using rabbit polyclonal anti- B . burgdorferi antibodies followed by goat anti-rabbit <t>Alexa</t> <t>Fluor</t> 488 antibodies to visualize spirochetes (green). Tick cells were visualized by staining with WGA-Alexa Fluor 594 (red). Scale bar = 10 μm. (B) Quantification of Borrelia in ticks that had fed on mice infected with wild-type (black circles), rel Bbu - (white circles) or rel Bbu - pBS- rel Bbu (gray circles) strains. Total DNA was isolated from larvae that had fed to repletion (fed larvae) or after larvae had molted to nymphs (unfed nymph) or one week after nymphs had fed to repletion on uninfected mice (fed nymph). The number of B . burgdorferi genome equivalents per tick was determined by qPCR using TaqMan primers/probe to flaB . The difference between the number of WT and rel Bbu - in fed nymphs was statistically significant ( P = 0.018) by a one-way ANOVA with a Tukey’s post hoc test.
    Goat Anti Rabbit Alexa Fluor 488 Antibodies, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit alexa fluor 488 antibodies/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Expression of integrin α 6 β 1 and stiffness of OPCs at different pH. ( a ) Expression level of integrin α 6 β 1 at different extracellular pH, evaluated by OPC immunostaining against α 6 β 1 (with Alexa Fluor-488 fluorochrome) and analysis of cell fluorescence using flow cytometry (BD LSR Fortessa). Data are geometric mean fluorescence intensities averaged over three experiments, each conducted in triplicate, and presented relative to value obtained for pH 7.0. No statistical difference was observed between any pH conditions. ( b ) Cell stiffness at pH 6.0 and 7.0, evaluated using AFM-enabled nanoindentation. Data are mean of Young's elastic modulus measured for 15 cells per condition. No statistical difference was observed between pH 6.0 and 7.0. Error bars are SEM. Colors correspond to cell media pH.

    Journal: PLoS ONE

    Article Title: Extracellular Acidic pH Inhibits Oligodendrocyte Precursor Viability, Migration, and Differentiation

    doi: 10.1371/journal.pone.0076048

    Figure Lengend Snippet: Expression of integrin α 6 β 1 and stiffness of OPCs at different pH. ( a ) Expression level of integrin α 6 β 1 at different extracellular pH, evaluated by OPC immunostaining against α 6 β 1 (with Alexa Fluor-488 fluorochrome) and analysis of cell fluorescence using flow cytometry (BD LSR Fortessa). Data are geometric mean fluorescence intensities averaged over three experiments, each conducted in triplicate, and presented relative to value obtained for pH 7.0. No statistical difference was observed between any pH conditions. ( b ) Cell stiffness at pH 6.0 and 7.0, evaluated using AFM-enabled nanoindentation. Data are mean of Young's elastic modulus measured for 15 cells per condition. No statistical difference was observed between pH 6.0 and 7.0. Error bars are SEM. Colors correspond to cell media pH.

    Article Snippet: Secondary antibodies included goat anti-mouse IgM Alexa Fluor 488 (Invitrogen), goat anti-rabbit IgG Alexa Fluor 488 (Invitrogen), and rabbit anti-rat IgG Alexa Fluor 488 (Invitrogen).

    Techniques: Expressing, Immunostaining, Fluorescence, Flow Cytometry, Cytometry

    The N1FR2011N-Vpu blocks anterograde transport of tetherin. ( A–D ) The nuclei of HT1080 cells, grown on coverslips, were co-microinjected with an expression plasmid encoding human tetherin with an internal HA tag together with vectors encoding either GFP ( A ) or C-terminally AU1 tagged Vpus: NL4-3 ( B ), N1FR2011 ( C ) and YBF30 ( D ). ∼200 cells were microinjected per plasmid combination. Subsequently, cells were cultivated for 1, 2 or 6 hours and then fixed, permeabilized and stained with an anti-HA mAb followed by an Alexa 568-conjugated secondary a (red staining) to detect newly synthesized tetherin together with an anti-AU1 rab followed by Alexa 488-conjugated secondary antibody (green staining) to detect newly synthesized Vpu proteins. Microphotographs shown are representative for three independent experiments. Cell circumferences are indicated. Scale bars: 10 µm. ( E ) Stainings were categorized into cells, which, besides intracellular staining, also displayed a clear plasma membrane staining (“plasma membrane”) or cells with an exclusively intracellular staining (“intracellular”). Histogram bars depict the relative percentage of cells for each time point from at least 150 cells that were analyzed out of three independent microinjection experiments.

    Journal: PLoS Pathogens

    Article Title: Human Tetherin Exerts Strong Selection Pressure on the HIV-1 Group N Vpu Protein

    doi: 10.1371/journal.ppat.1003093

    Figure Lengend Snippet: The N1FR2011N-Vpu blocks anterograde transport of tetherin. ( A–D ) The nuclei of HT1080 cells, grown on coverslips, were co-microinjected with an expression plasmid encoding human tetherin with an internal HA tag together with vectors encoding either GFP ( A ) or C-terminally AU1 tagged Vpus: NL4-3 ( B ), N1FR2011 ( C ) and YBF30 ( D ). ∼200 cells were microinjected per plasmid combination. Subsequently, cells were cultivated for 1, 2 or 6 hours and then fixed, permeabilized and stained with an anti-HA mAb followed by an Alexa 568-conjugated secondary a (red staining) to detect newly synthesized tetherin together with an anti-AU1 rab followed by Alexa 488-conjugated secondary antibody (green staining) to detect newly synthesized Vpu proteins. Microphotographs shown are representative for three independent experiments. Cell circumferences are indicated. Scale bars: 10 µm. ( E ) Stainings were categorized into cells, which, besides intracellular staining, also displayed a clear plasma membrane staining (“plasma membrane”) or cells with an exclusively intracellular staining (“intracellular”). Histogram bars depict the relative percentage of cells for each time point from at least 150 cells that were analyzed out of three independent microinjection experiments.

    Article Snippet: Newly synthesized Vpu molecules fused to AU-1 were detected using rabbit anti-AU1 (Covance) and anti-rabbit Alexa 488 secondary antibody (Invitrogen).

    Techniques: Expressing, Plasmid Preparation, Staining, Synthesized

    Photolyase activation prevents UVB-induced DNA damage and restores cell viability HaCaT keratinocytes (n = 2 × 10 5 cell) were treated with 20 or 40 mJ/cm 2 UVB, then cells were harvested. CPD formation from (a) genomic and (b) mitochondrial DNA was measured by CPD ELISA (n = 3) after UVB exposure, as indicated. (c) Cell viability was assessed by Annexin V-Alexa-488 and Propidium iodide labeling (n = 5) 24 after UVB exposure. (d) Cells were harvested 24 h post-irradiation and a clonogenic assay was performed (5000 cells/dish, n=4). (e) Cell cycle progression was assessed by Propidium Iodide incorporation (n = 4) 24 after UVB exposure. - represents where photolyase is inactive, + represents where photolyase is active *; **, and *** indicate statistically significant differences at p

    Journal: Redox Biology

    Article Title: Cyclobutane pyrimidine dimers from UVB exposure induce a hypermetabolic state in keratinocytes via mitochondrial oxidative stress

    doi: 10.1016/j.redox.2020.101808

    Figure Lengend Snippet: Photolyase activation prevents UVB-induced DNA damage and restores cell viability HaCaT keratinocytes (n = 2 × 10 5 cell) were treated with 20 or 40 mJ/cm 2 UVB, then cells were harvested. CPD formation from (a) genomic and (b) mitochondrial DNA was measured by CPD ELISA (n = 3) after UVB exposure, as indicated. (c) Cell viability was assessed by Annexin V-Alexa-488 and Propidium iodide labeling (n = 5) 24 after UVB exposure. (d) Cells were harvested 24 h post-irradiation and a clonogenic assay was performed (5000 cells/dish, n=4). (e) Cell cycle progression was assessed by Propidium Iodide incorporation (n = 4) 24 after UVB exposure. - represents where photolyase is inactive, + represents where photolyase is active *; **, and *** indicate statistically significant differences at p

    Article Snippet: The next day, slides were washed three times with PBS and incubated with Alexa-488 conjugated goat anti-rabbit antibody (Thermo Fisher Scientific) for K1 visualization.

    Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Labeling, Irradiation, Clonogenic Assay

    Tick persistence depends on rel Bbu . (A) Immunofluorescence microscopy of ticks allowed to feed on mice infected with wild-type (WT), rel Bbu mutant ( rel Bbu - ) or rel Bbu - pBS- rel Bbu strains. Ticks were dissected and fixed on slides one week after naïve larvae had fed to repletion (fed larvae) or after larvae had molted to nymphs (unfed nymph) or one week after nymphs had fed to repletion on uninfected mice (fed nymph). Samples were processed for IF microscopy using rabbit polyclonal anti- B . burgdorferi antibodies followed by goat anti-rabbit Alexa Fluor 488 antibodies to visualize spirochetes (green). Tick cells were visualized by staining with WGA-Alexa Fluor 594 (red). Scale bar = 10 μm. (B) Quantification of Borrelia in ticks that had fed on mice infected with wild-type (black circles), rel Bbu - (white circles) or rel Bbu - pBS- rel Bbu (gray circles) strains. Total DNA was isolated from larvae that had fed to repletion (fed larvae) or after larvae had molted to nymphs (unfed nymph) or one week after nymphs had fed to repletion on uninfected mice (fed nymph). The number of B . burgdorferi genome equivalents per tick was determined by qPCR using TaqMan primers/probe to flaB . The difference between the number of WT and rel Bbu - in fed nymphs was statistically significant ( P = 0.018) by a one-way ANOVA with a Tukey’s post hoc test.

    Journal: PLoS Pathogens

    Article Title: The Borrelia burgdorferi RelA/SpoT Homolog and Stringent Response Regulate Survival in the Tick Vector and Global Gene Expression during Starvation

    doi: 10.1371/journal.ppat.1005160

    Figure Lengend Snippet: Tick persistence depends on rel Bbu . (A) Immunofluorescence microscopy of ticks allowed to feed on mice infected with wild-type (WT), rel Bbu mutant ( rel Bbu - ) or rel Bbu - pBS- rel Bbu strains. Ticks were dissected and fixed on slides one week after naïve larvae had fed to repletion (fed larvae) or after larvae had molted to nymphs (unfed nymph) or one week after nymphs had fed to repletion on uninfected mice (fed nymph). Samples were processed for IF microscopy using rabbit polyclonal anti- B . burgdorferi antibodies followed by goat anti-rabbit Alexa Fluor 488 antibodies to visualize spirochetes (green). Tick cells were visualized by staining with WGA-Alexa Fluor 594 (red). Scale bar = 10 μm. (B) Quantification of Borrelia in ticks that had fed on mice infected with wild-type (black circles), rel Bbu - (white circles) or rel Bbu - pBS- rel Bbu (gray circles) strains. Total DNA was isolated from larvae that had fed to repletion (fed larvae) or after larvae had molted to nymphs (unfed nymph) or one week after nymphs had fed to repletion on uninfected mice (fed nymph). The number of B . burgdorferi genome equivalents per tick was determined by qPCR using TaqMan primers/probe to flaB . The difference between the number of WT and rel Bbu - in fed nymphs was statistically significant ( P = 0.018) by a one-way ANOVA with a Tukey’s post hoc test.

    Article Snippet: Slides were washed 3 × 10 min in wash buffer (dPBS + 5 mM MgCl2 + 1% goat serum (Gibco, Life Technologies)) and then incubated with rabbit polyclonal anti-B . burgdorferi antibodies (a gift from Tom Schwan) at 1:200 dilution for 1 h. Slides were washed again as above and the primary antibodies detected using goat anti-rabbit Alexa Fluor 488 antibodies (Molecular Probes) at 1:500 dilution for 1 h. Slides were washed 2 × 10 min in wash buffer.

    Techniques: Immunofluorescence, Microscopy, Mouse Assay, Infection, Mutagenesis, Staining, Whole Genome Amplification, Isolation, Real-time Polymerase Chain Reaction