goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate  (Millipore)


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    Structured Review

    Millipore goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate
    Virion-specific <t>IgG</t> and neutralizing-antibody responses in the DNA primed/FI-MCMV-boosted or live virus-vaccinated mice. Mice from each group vaccinated with either pc3-Ua plus PBS plus alum (two mice per group), All-U pDNA plus FI+alum (six mice
    Goat Anti Mouse Immunoglobulin G Igg Horseradish Peroxidase Conjugate, supplied by Millipore, used in various techniques. Bioz Stars score: 82/100, based on 354 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate/product/Millipore
    Average 82 stars, based on 354 article reviews
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    goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate - by Bioz Stars, 2020-02
    82/100 stars

    Images

    1) Product Images from "Systemic Priming-Boosting Immunization with a Trivalent Plasmid DNA and Inactivated Murine Cytomegalovirus (MCMV) Vaccine Provides Long-Term Protection against Viral Replication following Systemic or Mucosal MCMV Challenge"

    Article Title: Systemic Priming-Boosting Immunization with a Trivalent Plasmid DNA and Inactivated Murine Cytomegalovirus (MCMV) Vaccine Provides Long-Term Protection against Viral Replication following Systemic or Mucosal MCMV Challenge

    Journal:

    doi: 10.1128/JVI.79.1.159-175.2005

    Virion-specific IgG and neutralizing-antibody responses in the DNA primed/FI-MCMV-boosted or live virus-vaccinated mice. Mice from each group vaccinated with either pc3-Ua plus PBS plus alum (two mice per group), All-U pDNA plus FI+alum (six mice
    Figure Legend Snippet: Virion-specific IgG and neutralizing-antibody responses in the DNA primed/FI-MCMV-boosted or live virus-vaccinated mice. Mice from each group vaccinated with either pc3-Ua plus PBS plus alum (two mice per group), All-U pDNA plus FI+alum (six mice

    Techniques Used: Mouse Assay

    Virion-specific IgG and neutralizing-antibody responses in vaccinated mice. On the weeks of the experiment shown in Fig. , four to eight mice per vaccine group were retroorbitally bled and sera were prepared. Arrows and numbers indicate
    Figure Legend Snippet: Virion-specific IgG and neutralizing-antibody responses in vaccinated mice. On the weeks of the experiment shown in Fig. , four to eight mice per vaccine group were retroorbitally bled and sera were prepared. Arrows and numbers indicate

    Techniques Used: Mouse Assay

    2) Product Images from "Integrin α2β1 Expression Regulates Matrix Metalloproteinase-1-Dependent Bronchial Epithelial Repair in Pulmonary Tuberculosis"

    Article Title: Integrin α2β1 Expression Regulates Matrix Metalloproteinase-1-Dependent Bronchial Epithelial Repair in Pulmonary Tuberculosis

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.01348

    Matrix type I collagen induces α2β1 integrin clustering and actin colocalization without affecting integrin surface expression. Human bronchial epithelial cells (HBECs) were cultured in the in the presence of matrix or soluble Coll-I or without extracellular matrix (ECM) as controls and stimulated with CoMtb or control medium. (A) Confocal microscopy figures of CoMtb-stimulated HBECs. Cells were stained with DAPI (blue) for nucleic acids, phalloidin conjugated with Alexa Fluor 594 (red) for F-actin, and FITC-conjugated anti-integrin α2 antibodies (green) for integrin subunit α2. Yellow color shows co-localization of F-actin and integrin α2β1 in merged images. Scale bar: 50 µm. (B,C) Control and CoMtb-stimulated HBECs were labeled with FITC-conjugated anti-integrin α2 or α3 and analyzed by FACS. FITC-conjugated mouse IgG1 was used as isotype control. Plots show mean fluorescence intensities (MFI) and SD for (B) α2 and (C) α3 integrins of three independent experiments.
    Figure Legend Snippet: Matrix type I collagen induces α2β1 integrin clustering and actin colocalization without affecting integrin surface expression. Human bronchial epithelial cells (HBECs) were cultured in the in the presence of matrix or soluble Coll-I or without extracellular matrix (ECM) as controls and stimulated with CoMtb or control medium. (A) Confocal microscopy figures of CoMtb-stimulated HBECs. Cells were stained with DAPI (blue) for nucleic acids, phalloidin conjugated with Alexa Fluor 594 (red) for F-actin, and FITC-conjugated anti-integrin α2 antibodies (green) for integrin subunit α2. Yellow color shows co-localization of F-actin and integrin α2β1 in merged images. Scale bar: 50 µm. (B,C) Control and CoMtb-stimulated HBECs were labeled with FITC-conjugated anti-integrin α2 or α3 and analyzed by FACS. FITC-conjugated mouse IgG1 was used as isotype control. Plots show mean fluorescence intensities (MFI) and SD for (B) α2 and (C) α3 integrins of three independent experiments.

    Techniques Used: Expressing, Cell Culture, Confocal Microscopy, Staining, Labeling, FACS, Fluorescence

    3) Product Images from "Disruption of the serine/threonine protein kinase H affects phthiocerol dimycocerosates synthesis in Mycobacterium tuberculosis"

    Article Title: Disruption of the serine/threonine protein kinase H affects phthiocerol dimycocerosates synthesis in Mycobacterium tuberculosis

    Journal: Microbiology

    doi: 10.1099/mic.0.062067-0

    (a) Immunofluorescence microscopy analysis. Bacteria were labelled with rhodamine, and scFv antibodies against PDIMs were used as primary antibodies. Anti-MBP antibody coupled to goat anti-mouse IgG-FITC was used as the secondary antibody. The merged images are shown in the panels on the right. M. smegmatis was used as a negative control. (b) Immunofluorescence detection. Data represent the means± sd of green fluorescence intensity (labelled PDIMs) in arbitrary units (a.u.), which corresponds to labelled PDIMs, per cellular area; n = 50 single bacterial cells. * P ≤0.05.
    Figure Legend Snippet: (a) Immunofluorescence microscopy analysis. Bacteria were labelled with rhodamine, and scFv antibodies against PDIMs were used as primary antibodies. Anti-MBP antibody coupled to goat anti-mouse IgG-FITC was used as the secondary antibody. The merged images are shown in the panels on the right. M. smegmatis was used as a negative control. (b) Immunofluorescence detection. Data represent the means± sd of green fluorescence intensity (labelled PDIMs) in arbitrary units (a.u.), which corresponds to labelled PDIMs, per cellular area; n = 50 single bacterial cells. * P ≤0.05.

    Techniques Used: Immunofluorescence, Microscopy, Negative Control, Fluorescence

    4) Product Images from "Dopamine-Dependent Tuning of Striatal Inhibitory Synaptogenesis"

    Article Title: Dopamine-Dependent Tuning of Striatal Inhibitory Synaptogenesis

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.4411-09.2010

    Dopamine receptor activation decreases the cell-surface expression of GABA A receptors via a dynamin-dependent pathway. A , B , Internalization of GABA A receptors was visualized by staining with β 2/3 -specific antibody in the presence of vehicle, SKF-38393 ( A ), or quinpirole ( B ), as described in Materials and Methods. Binding of β 2/3 -specific antibody to the surface expressed GABA A receptors was visualized using a secondary anti-mouse IgG coupled to Alexa555 (in red), while binding of the same antibody to internalized receptors was visualized using a secondary anti-mouse IgG coupled to Alexa488 (in green). Scale bars correspond to 10 μm. C , The decrease in surface β 2/3 subunit levels caused by SKF-38393 (SKF) was significantly attenuated in the presence of a dynamin inhibitory peptide (SKF/DynP), but not in the presence of its scrambled control (SKF/DynPC). D , The decrease in surface β 2/3 subunit levels caused by quinpirole (Q) was also significantly attenuated in the presence of a dynamin inhibitory peptide (Q/DynP), but not by its scrambled control (Q/DynPC). Bars represent mean ± SEM. Statistical analysis was performed using one-way ANOVA with Dunnett post hoc analysis versus control (* p
    Figure Legend Snippet: Dopamine receptor activation decreases the cell-surface expression of GABA A receptors via a dynamin-dependent pathway. A , B , Internalization of GABA A receptors was visualized by staining with β 2/3 -specific antibody in the presence of vehicle, SKF-38393 ( A ), or quinpirole ( B ), as described in Materials and Methods. Binding of β 2/3 -specific antibody to the surface expressed GABA A receptors was visualized using a secondary anti-mouse IgG coupled to Alexa555 (in red), while binding of the same antibody to internalized receptors was visualized using a secondary anti-mouse IgG coupled to Alexa488 (in green). Scale bars correspond to 10 μm. C , The decrease in surface β 2/3 subunit levels caused by SKF-38393 (SKF) was significantly attenuated in the presence of a dynamin inhibitory peptide (SKF/DynP), but not in the presence of its scrambled control (SKF/DynPC). D , The decrease in surface β 2/3 subunit levels caused by quinpirole (Q) was also significantly attenuated in the presence of a dynamin inhibitory peptide (Q/DynP), but not by its scrambled control (Q/DynPC). Bars represent mean ± SEM. Statistical analysis was performed using one-way ANOVA with Dunnett post hoc analysis versus control (* p

    Techniques Used: Activation Assay, Expressing, Staining, Binding Assay

    5) Product Images from "Basolateral Targeting of ERBB2 Is Dependent on a Novel Bipartite Juxtamembrane Sorting Signal but Independent of the C-Terminal ERBIN-Binding Domain"

    Article Title: Basolateral Targeting of ERBB2 Is Dependent on a Novel Bipartite Juxtamembrane Sorting Signal but Independent of the C-Terminal ERBIN-Binding Domain

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.22.18.6553-6563.2002

    ERBB2 contains a juxtamembrane basolateral sorting signal. (A) MDCK cells containing an inducible ERBB2 expression vector were grown on transwell filters to confluence before overnight induction with tetracycline (1 μg/ml). After induction, cells were fixed in methanol, incubated with a rat antibody to ZO-1 and a mouse monoclonal antibody against ERBB2, and subsequently stained with FITC-conjugated goat anti-mouse IgG and TRITC-conjugated goat anti-rat IgG. Panels show confocal images of the XY sections through the tight junctions, and XZ sections with the apical membrane uppermost. (B) Immunoblot of an MDCK cell clone showing induced and uninduced ERBB2 expression. tet, tetracycline. (C) Schematic depiction of ERBB2 vectors showing the transmembrane domain (TM) and the extent of the C-terminal truncations in the juxtamembrane domain (amino acids 680 to 710). The asterisk shows the position of the introduced termination codon. (D) Immunoblot analysis of truncation mutants analyzed by confocal microscopy in panel E. (E) Confocal microscopy images of XY sections and XZ sections prepared and stained as for panel A. Bars, 5 μm.
    Figure Legend Snippet: ERBB2 contains a juxtamembrane basolateral sorting signal. (A) MDCK cells containing an inducible ERBB2 expression vector were grown on transwell filters to confluence before overnight induction with tetracycline (1 μg/ml). After induction, cells were fixed in methanol, incubated with a rat antibody to ZO-1 and a mouse monoclonal antibody against ERBB2, and subsequently stained with FITC-conjugated goat anti-mouse IgG and TRITC-conjugated goat anti-rat IgG. Panels show confocal images of the XY sections through the tight junctions, and XZ sections with the apical membrane uppermost. (B) Immunoblot of an MDCK cell clone showing induced and uninduced ERBB2 expression. tet, tetracycline. (C) Schematic depiction of ERBB2 vectors showing the transmembrane domain (TM) and the extent of the C-terminal truncations in the juxtamembrane domain (amino acids 680 to 710). The asterisk shows the position of the introduced termination codon. (D) Immunoblot analysis of truncation mutants analyzed by confocal microscopy in panel E. (E) Confocal microscopy images of XY sections and XZ sections prepared and stained as for panel A. Bars, 5 μm.

    Techniques Used: Expressing, Plasmid Preparation, Incubation, Staining, Confocal Microscopy

    6) Product Images from "Characterization of Novel PI3K? Inhibitors as Potential Therapeutics for SLE and Lupus Nephritis in Pre-Clinical Studies"

    Article Title: Characterization of Novel PI3K? Inhibitors as Potential Therapeutics for SLE and Lupus Nephritis in Pre-Clinical Studies

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2014.00233

    MSC2360844 abrogates lupus nephritis development in accelerated NZB/W F1 SLE model . (A) Incidence of proteinuria and (B) mean UPCR overtime in mice treated once daily with MSC2360844 at 6.6, 22, or 66 mg/kg, or MMF at 300 mg/kg with an early dosing regimen, or MSC2360844 at 66 mg/kg with a late dosing regimen (L). (C) UPCR over time in mice treated with BAFF-R-Ig or mFc as control. (D) Kidney sections obtained at the end of experiment were stained with H E (upper panel) or immunofluorescence (lower panel) with anti-IgG2a FITC Ab. Magnification, 200×. (E) Histopathological scores in (D) (upper panel). Each dot represents one mouse. Bar represents mean value. (F) ASCs in the spleen measured with ELISpot at the end of experiment. Each dot represents one mouse. Bar represents mean value. (G) Levels of anti-dsDNA IgG at the end of experiment. Each dot represents one mouse. Bar represents mean value. (H) Ex vivo anti-IgD-induced CD69 up-regulation on B220 + cells in whole blood 1 h post last dosing after 2 week of MSC2360844 treatment. Each dot represents one mouse. Bar represents mean value. (I) Relationship between PD marker pAkt and UPCR. The pAkt PK/PD model was used to mimic pAkt response under efficacy experiment conditions. * p
    Figure Legend Snippet: MSC2360844 abrogates lupus nephritis development in accelerated NZB/W F1 SLE model . (A) Incidence of proteinuria and (B) mean UPCR overtime in mice treated once daily with MSC2360844 at 6.6, 22, or 66 mg/kg, or MMF at 300 mg/kg with an early dosing regimen, or MSC2360844 at 66 mg/kg with a late dosing regimen (L). (C) UPCR over time in mice treated with BAFF-R-Ig or mFc as control. (D) Kidney sections obtained at the end of experiment were stained with H E (upper panel) or immunofluorescence (lower panel) with anti-IgG2a FITC Ab. Magnification, 200×. (E) Histopathological scores in (D) (upper panel). Each dot represents one mouse. Bar represents mean value. (F) ASCs in the spleen measured with ELISpot at the end of experiment. Each dot represents one mouse. Bar represents mean value. (G) Levels of anti-dsDNA IgG at the end of experiment. Each dot represents one mouse. Bar represents mean value. (H) Ex vivo anti-IgD-induced CD69 up-regulation on B220 + cells in whole blood 1 h post last dosing after 2 week of MSC2360844 treatment. Each dot represents one mouse. Bar represents mean value. (I) Relationship between PD marker pAkt and UPCR. The pAkt PK/PD model was used to mimic pAkt response under efficacy experiment conditions. * p

    Techniques Used: Mouse Assay, Staining, Immunofluorescence, Enzyme-linked Immunospot, Ex Vivo, Marker

    7) Product Images from "A mouse model of systemic lupus erythematosus responds better to soluble TACI than to soluble BAFFR, correlating with depletion of plasma cells"

    Article Title: A mouse model of systemic lupus erythematosus responds better to soluble TACI than to soluble BAFFR, correlating with depletion of plasma cells

    Journal: European Journal of Immunology

    doi: 10.1002/eji.201746934

    mTACI‐Fc is more effective than mBAFFR‐Fc in diminishing the bone marrow PC compartment and in preventing autoantibody increase and renal immunopathology in NZB/NZW F1 mice. (A). Antibody secreting cells (ASC) retrieved from spleens of 37‐week‐old NZB/NZW F1 mice subjected to the indicated treatments, as determined by ELISPOT. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 25/15/11/4. Values were obtained from titrations performed in triplicates. (B). Numbers of PCs (CD138 + ) derived from spleens of NZB/NZW F1 mice. Absolute cell numbers were determined by FACS from specimens taken at termination of the experiment and included analyses of animals that had been sacrificed at earlier times throughout the duration of the study. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 44/15/15/14. (C). Same as panel A, but for the bone marrow of 37‐week‐old mice. (D). Same as panel B, but for absolute numbers of CD138 + cells in the bone marrow of two femurs and for 37‐week‐old mice only. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 25/15/11/4. (E). Evolution of anti‐dsDNA IgG levels in groups of control and treated mice. Mice that had to be sacrificed before week 37 are indicated with black circles. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 44/15/15/14. (F). Log 10 of fold change of anti‐dsDNA levels between week 24 and week 37. Only NZB/NZW F1 mice with less than 100 arbitrary units of anti‐dsDNA antibody at week 24 were included in this analysis. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 28/10/12/8. (G). Renal histology scores for the different treatment groups at sacrifice (week 37 or earlier). n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 45/15/15/15. (H). Relationship between anti‐dsDNA titers at early time points (average of titers measured at weeks 24 and 28) and a proteinuria score taking into account timing of apparition and severity of proteinuria. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 43/15/15/14. Panels A‐H are from an experiment that was performed once. Panels A‐D, F, and G show mean of each group ± SEM. Symbols represent individual mice. Statistical analysis was performed with one‐way ANOVA followed by Bonferroni comparing controls to each treatment. ns: non‐significant; * p
    Figure Legend Snippet: mTACI‐Fc is more effective than mBAFFR‐Fc in diminishing the bone marrow PC compartment and in preventing autoantibody increase and renal immunopathology in NZB/NZW F1 mice. (A). Antibody secreting cells (ASC) retrieved from spleens of 37‐week‐old NZB/NZW F1 mice subjected to the indicated treatments, as determined by ELISPOT. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 25/15/11/4. Values were obtained from titrations performed in triplicates. (B). Numbers of PCs (CD138 + ) derived from spleens of NZB/NZW F1 mice. Absolute cell numbers were determined by FACS from specimens taken at termination of the experiment and included analyses of animals that had been sacrificed at earlier times throughout the duration of the study. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 44/15/15/14. (C). Same as panel A, but for the bone marrow of 37‐week‐old mice. (D). Same as panel B, but for absolute numbers of CD138 + cells in the bone marrow of two femurs and for 37‐week‐old mice only. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 25/15/11/4. (E). Evolution of anti‐dsDNA IgG levels in groups of control and treated mice. Mice that had to be sacrificed before week 37 are indicated with black circles. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 44/15/15/14. (F). Log 10 of fold change of anti‐dsDNA levels between week 24 and week 37. Only NZB/NZW F1 mice with less than 100 arbitrary units of anti‐dsDNA antibody at week 24 were included in this analysis. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 28/10/12/8. (G). Renal histology scores for the different treatment groups at sacrifice (week 37 or earlier). n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 45/15/15/15. (H). Relationship between anti‐dsDNA titers at early time points (average of titers measured at weeks 24 and 28) and a proteinuria score taking into account timing of apparition and severity of proteinuria. n for controls/TACI‐Fc/BAFFR‐Fc/Apry‐1‐1 were 43/15/15/14. Panels A‐H are from an experiment that was performed once. Panels A‐D, F, and G show mean of each group ± SEM. Symbols represent individual mice. Statistical analysis was performed with one‐way ANOVA followed by Bonferroni comparing controls to each treatment. ns: non‐significant; * p

    Techniques Used: Mouse Assay, Enzyme-linked Immunospot, Derivative Assay, FACS

    m and hTACI‐Fc, not mBAFFR‐Fc, efficiently deplete bone marrow PC in immunized C56BL/6 mice, while efficacy of Apry‐1‐1 alone or with mBAFFR‐Fc varies as PCs mature. C57BL/6 mice were immunized at 8 weeks of age with NP‐conjugated keyhole limpet hemocyanin precipitated in Alum. 2, 5, or 12 weeks later, mice were treated for two additional weeks, three times a week, with either mTACI‐Fc, hTACI‐Fc, mBAFFR‐Fc, anti‐mAPRIL Apry‐1‐1 or control reagents. Data shown combine results of two independent experiments each with n = 5 animals per group (with the exception of hTACI‐Fc and intravenous immunoglobulin (IVIG) that were performed once). (A). Quantification of mouse BAFF‐blocking activity in sera of treated mice at the end of the experiment, using a BCMA:Fas reporter cell line. (B). Same as panel C, but for the quantification of mouse APRIL‐blocking activity. (C). Relative absolute numbers of splenic B cells (CD19 + /B220 + ), compared to the mean of a control group (mIg + mFc). (D). Relative absolute numbers of bone marrow NP‐specific IgG antibody‐secreting cells (ASC) measured by ELISPOT compared to the mean of a control group. Mice were treated with the indicated inhibitors from weeks 2 to 4 (weeks 2–4), 5 to 7 (weeks 5–7), or 12 to 14 (weeks 12–14) post immunization and analyzed at the end of this 2‐week treatment. All graphs show mean ± SEM. Symbols represent individual mice. Analyses in panels A and B were performed twice with similar results. Statistical analysis was performed with one way ANOVA followed by Bonferroni comparing controls to each treatment (A–D). ns: nonsignificant; * p
    Figure Legend Snippet: m and hTACI‐Fc, not mBAFFR‐Fc, efficiently deplete bone marrow PC in immunized C56BL/6 mice, while efficacy of Apry‐1‐1 alone or with mBAFFR‐Fc varies as PCs mature. C57BL/6 mice were immunized at 8 weeks of age with NP‐conjugated keyhole limpet hemocyanin precipitated in Alum. 2, 5, or 12 weeks later, mice were treated for two additional weeks, three times a week, with either mTACI‐Fc, hTACI‐Fc, mBAFFR‐Fc, anti‐mAPRIL Apry‐1‐1 or control reagents. Data shown combine results of two independent experiments each with n = 5 animals per group (with the exception of hTACI‐Fc and intravenous immunoglobulin (IVIG) that were performed once). (A). Quantification of mouse BAFF‐blocking activity in sera of treated mice at the end of the experiment, using a BCMA:Fas reporter cell line. (B). Same as panel C, but for the quantification of mouse APRIL‐blocking activity. (C). Relative absolute numbers of splenic B cells (CD19 + /B220 + ), compared to the mean of a control group (mIg + mFc). (D). Relative absolute numbers of bone marrow NP‐specific IgG antibody‐secreting cells (ASC) measured by ELISPOT compared to the mean of a control group. Mice were treated with the indicated inhibitors from weeks 2 to 4 (weeks 2–4), 5 to 7 (weeks 5–7), or 12 to 14 (weeks 12–14) post immunization and analyzed at the end of this 2‐week treatment. All graphs show mean ± SEM. Symbols represent individual mice. Analyses in panels A and B were performed twice with similar results. Statistical analysis was performed with one way ANOVA followed by Bonferroni comparing controls to each treatment (A–D). ns: nonsignificant; * p

    Techniques Used: Mouse Assay, Blocking Assay, Activity Assay, Enzyme-linked Immunospot

    8) Product Images from "Mesenchymal stem cell transplantation can restore lupus disease-associated miRNA expression and Th1/Th2 ratios in a murine model of SLE"

    Article Title: Mesenchymal stem cell transplantation can restore lupus disease-associated miRNA expression and Th1/Th2 ratios in a murine model of SLE

    Journal: Scientific Reports

    doi: 10.1038/srep38237

    Histological staining and immunofluorescence of kidney samples. ( A ) Periodic acid-Schiff staining of kidney tissue. Bar = 50 μm. ( B ) Immunofluorescent staining of kidney samples from the experimental groups was conducted using FITC-anti IgG or FITC-anti C3 antibodies to compare the effects of various treatments. Bar = 50 μm. ( C ) IgG and C3 deposition scores in the glomerula. The intensity of fluorescence was graded on a scale of 0 (none) to 4 (great fluorescence intensity). Data obtained from each group were compared using ANOVA followed by post hoc Tukey’s multiple comparison tests (C: n = 13, Y: n = 15, H: n = 15, and N: n = 10). * significant ( p
    Figure Legend Snippet: Histological staining and immunofluorescence of kidney samples. ( A ) Periodic acid-Schiff staining of kidney tissue. Bar = 50 μm. ( B ) Immunofluorescent staining of kidney samples from the experimental groups was conducted using FITC-anti IgG or FITC-anti C3 antibodies to compare the effects of various treatments. Bar = 50 μm. ( C ) IgG and C3 deposition scores in the glomerula. The intensity of fluorescence was graded on a scale of 0 (none) to 4 (great fluorescence intensity). Data obtained from each group were compared using ANOVA followed by post hoc Tukey’s multiple comparison tests (C: n = 13, Y: n = 15, H: n = 15, and N: n = 10). * significant ( p

    Techniques Used: Staining, Immunofluorescence, Fluorescence

    9) Product Images from "Temporal regulation of Lsp1 O-GlcNAcylation and phosphorylation during apoptosis of activated B cells"

    Article Title: Temporal regulation of Lsp1 O-GlcNAcylation and phosphorylation during apoptosis of activated B cells

    Journal: Nature Communications

    doi: 10.1038/ncomms12526

    Dynamic interplay between O -GlcNAcylation and phosphorylation on Lsp1 in B cells after anti-IgM stimulation . ( a ) Mass spectrometric results revealing 12 mapped phosphorylation sites and 1 O -GlcNAcylation site on Lsp1. ( b ) Mapping of the O -GlcNAcylation site 208-KSQPTLPISTIDER-221 of Lsp1 using ETD fragmentation during MS/MS analysis. The ions, c 1 + (146.3 Da), c 2 + (436.3 Da), z+1 12 + (1353.5 Da), z+1 13 + (1643.8 Da) suggest that S209 is O -GlcNAcylated. ( c ) Lysates prepared from Ramos B cells overexpressing the vector control, Flag-EGFP-tagged WT or S209A Lsp1 were subjected to a pull-down assay using sWGA agarose beads, followed by immunoblotting (IB) with an anti-Flag antibody. ( d ) Lysates from 293T cells ectopically expressing OGT and either vector, Flag-EGFP-tagged WT or S209A Lsp1, were used for immunoprecipitation (IP) with anti-Flag, followed by IB with the indicated antibodies. ( e ) IB showing the levels of Lsp1, S243 phosphorylated Lsp1 and O -GlcNAcylated Lsp1 in anti-IgM (10 μg ml −1 ) stimulated mouse splenic B cells at indicated time points in an IP assay with control rabbit IgG (rIgG) or anti-Lsp1-specific antibody crosslinked to protein A agarose. The percentage of O -GlcNAcylated Lsp1 is indicated. ( f ) Sorted EGFP + Ramos B cells overexpressing Flag-EGFP-tagged WT or S209A Lsp1 were stimulated with anti-human IgM (25 μg ml −1 ) for 30 min, followed by IB analysis of Lsp1 S243 phosphorylation. One representative experiment out of three is shown. The relative levels of Lsp1 with phosphorylation at S243 were indicated. Results represent the mean±s.e.m. ( n =3). *** P
    Figure Legend Snippet: Dynamic interplay between O -GlcNAcylation and phosphorylation on Lsp1 in B cells after anti-IgM stimulation . ( a ) Mass spectrometric results revealing 12 mapped phosphorylation sites and 1 O -GlcNAcylation site on Lsp1. ( b ) Mapping of the O -GlcNAcylation site 208-KSQPTLPISTIDER-221 of Lsp1 using ETD fragmentation during MS/MS analysis. The ions, c 1 + (146.3 Da), c 2 + (436.3 Da), z+1 12 + (1353.5 Da), z+1 13 + (1643.8 Da) suggest that S209 is O -GlcNAcylated. ( c ) Lysates prepared from Ramos B cells overexpressing the vector control, Flag-EGFP-tagged WT or S209A Lsp1 were subjected to a pull-down assay using sWGA agarose beads, followed by immunoblotting (IB) with an anti-Flag antibody. ( d ) Lysates from 293T cells ectopically expressing OGT and either vector, Flag-EGFP-tagged WT or S209A Lsp1, were used for immunoprecipitation (IP) with anti-Flag, followed by IB with the indicated antibodies. ( e ) IB showing the levels of Lsp1, S243 phosphorylated Lsp1 and O -GlcNAcylated Lsp1 in anti-IgM (10 μg ml −1 ) stimulated mouse splenic B cells at indicated time points in an IP assay with control rabbit IgG (rIgG) or anti-Lsp1-specific antibody crosslinked to protein A agarose. The percentage of O -GlcNAcylated Lsp1 is indicated. ( f ) Sorted EGFP + Ramos B cells overexpressing Flag-EGFP-tagged WT or S209A Lsp1 were stimulated with anti-human IgM (25 μg ml −1 ) for 30 min, followed by IB analysis of Lsp1 S243 phosphorylation. One representative experiment out of three is shown. The relative levels of Lsp1 with phosphorylation at S243 were indicated. Results represent the mean±s.e.m. ( n =3). *** P

    Techniques Used: Mass Spectrometry, Plasmid Preparation, Pull Down Assay, Expressing, Immunoprecipitation

    10) Product Images from "Mosquito salivary allergen Aed a 3: cloning, comprehensive molecular analysis, and clinical evaluation"

    Article Title: Mosquito salivary allergen Aed a 3: cloning, comprehensive molecular analysis, and clinical evaluation

    Journal: Allergy

    doi: 10.1111/all.12812

    Fusion protein-selected antibodies recognize a 30 kDa Ae. aegypti salivary protein. A. Mouse antibodies. Ae. aegypti saliva proteins were separated by SDS-PAGE and immunoblotted with mouse anti-saliva serum (lane 1), fusion protein-selected mouse antibodies (lane 2), or PBST (lane 3), followed by incubation with peroxidase-conjugated goat anti-mouse IgG. B. Human antibodies. Ae. aegypti salivary proteins were separated by SDS-PAGE and immunoblotted with a pooled human mosquito-allergic serum (lane 1), fusion protein-selected human antibodies (lane 2), or PBST (lane 3), followed by incubation with mAb anti-human IgE.
    Figure Legend Snippet: Fusion protein-selected antibodies recognize a 30 kDa Ae. aegypti salivary protein. A. Mouse antibodies. Ae. aegypti saliva proteins were separated by SDS-PAGE and immunoblotted with mouse anti-saliva serum (lane 1), fusion protein-selected mouse antibodies (lane 2), or PBST (lane 3), followed by incubation with peroxidase-conjugated goat anti-mouse IgG. B. Human antibodies. Ae. aegypti salivary proteins were separated by SDS-PAGE and immunoblotted with a pooled human mosquito-allergic serum (lane 1), fusion protein-selected human antibodies (lane 2), or PBST (lane 3), followed by incubation with mAb anti-human IgE.

    Techniques Used: SDS Page, Incubation

    11) Product Images from "Disruption of the serine/threonine protein kinase H affects phthiocerol dimycocerosates synthesis in Mycobacterium tuberculosis"

    Article Title: Disruption of the serine/threonine protein kinase H affects phthiocerol dimycocerosates synthesis in Mycobacterium tuberculosis

    Journal: Microbiology

    doi: 10.1099/mic.0.062067-0

    (a) Immunofluorescence microscopy analysis. Bacteria were labelled with rhodamine, and scFv antibodies against PDIMs were used as primary antibodies. Anti-MBP antibody coupled to goat anti-mouse IgG-FITC was used as the secondary antibody. The merged images are shown in the panels on the right. M. smegmatis was used as a negative control. (b) Immunofluorescence detection. Data represent the means± sd of green fluorescence intensity (labelled PDIMs) in arbitrary units (a.u.), which corresponds to labelled PDIMs, per cellular area; n = 50 single bacterial cells. * P ≤0.05.
    Figure Legend Snippet: (a) Immunofluorescence microscopy analysis. Bacteria were labelled with rhodamine, and scFv antibodies against PDIMs were used as primary antibodies. Anti-MBP antibody coupled to goat anti-mouse IgG-FITC was used as the secondary antibody. The merged images are shown in the panels on the right. M. smegmatis was used as a negative control. (b) Immunofluorescence detection. Data represent the means± sd of green fluorescence intensity (labelled PDIMs) in arbitrary units (a.u.), which corresponds to labelled PDIMs, per cellular area; n = 50 single bacterial cells. * P ≤0.05.

    Techniques Used: Immunofluorescence, Microscopy, Negative Control, Fluorescence

    12) Product Images from "Selective Expression of CYP2A13 in Human Pancreatic ?-Islet Cells"

    Article Title: Selective Expression of CYP2A13 in Human Pancreatic ?-Islet Cells

    Journal: Drug Metabolism and Disposition

    doi: 10.1124/dmd.112.046359

    Immunofluorescent double staining images of CYP2A13, proinsulin C-peptide, and glucagon in adult human pancreatic islets (magnification, 400×). A-1 and B-1, rabbit anti-CYP2A13 antibody and FITC-conjugated goat anti-rabbit IgG were used as the
    Figure Legend Snippet: Immunofluorescent double staining images of CYP2A13, proinsulin C-peptide, and glucagon in adult human pancreatic islets (magnification, 400×). A-1 and B-1, rabbit anti-CYP2A13 antibody and FITC-conjugated goat anti-rabbit IgG were used as the

    Techniques Used: Double Staining

    13) Product Images from "Rapid Generation of Human-Like Neutralizing Monoclonal Antibodies in Urgent Preparedness for Influenza Pandemics and Virulent Infectious Diseases"

    Article Title: Rapid Generation of Human-Like Neutralizing Monoclonal Antibodies in Urgent Preparedness for Influenza Pandemics and Virulent Infectious Diseases

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0066276

    Analysis of lungs from mice treated with mAb 4E6 and 1H10 after infection with H5N1 virus A/Vietnam/1194/04. (A) Viral copy number in the lungs at day 6 post-infection was determined using Q-PCR. The results are expressed as the mean ± SEM. (B) Virus titer in the lungs at day 6 post-infection was measured using a plaque formation assay in MDCK cells. The results are expressed as the mean ± SEM. (C) Histopathology in pulmonary tissue (40x magnification). 1) Mouse treated with mAb 4E6 at 24 hours post-infection. 2) Mouse treated with mAb 1H10 at 24 hours post-infection. 3) Mice injected with an irrelevant IgG at 24 hours post-infection were used as the control. HE: H E staining. IHC: immunohistochemical staining using a mAb against influenza nucleoprotein.
    Figure Legend Snippet: Analysis of lungs from mice treated with mAb 4E6 and 1H10 after infection with H5N1 virus A/Vietnam/1194/04. (A) Viral copy number in the lungs at day 6 post-infection was determined using Q-PCR. The results are expressed as the mean ± SEM. (B) Virus titer in the lungs at day 6 post-infection was measured using a plaque formation assay in MDCK cells. The results are expressed as the mean ± SEM. (C) Histopathology in pulmonary tissue (40x magnification). 1) Mouse treated with mAb 4E6 at 24 hours post-infection. 2) Mouse treated with mAb 1H10 at 24 hours post-infection. 3) Mice injected with an irrelevant IgG at 24 hours post-infection were used as the control. HE: H E staining. IHC: immunohistochemical staining using a mAb against influenza nucleoprotein.

    Techniques Used: Mouse Assay, Infection, Polymerase Chain Reaction, Plaque Formation Assay, Histopathology, Injection, Staining, Immunohistochemistry

    Therapeutic efficacy of mAb 4E6 and 1H10 in a mouse model of H5N1 virus A/Vietnam/1194/04 infection. (A) The Kaplan-Meier survival curve. BALB/c mice (n=7 per group in two separate experiments) were infected intranasally with 10 LD 50 of A/Vietnam/1194/04 and at 24, 48, or 72 hours, were treated with a single intraperitoneal injection of mAb 4E6 or 1H10 at 10 mg/kg body weight. The control group received IgG at 10 mg/kg body weight at 24 hours post-infection. (B) Percentage weight change in mice treated with mAb 4E6 or 1H10 at different time points post-infection. The mice (n=7) were infected via intranasal inoculation with 10 LD50 of A/Vietnam/1203/4, followed by a single intraperitoneal injection of mAbs at 24, 48, or 72 hours post-infection, and their weights were monitored for 14 days. The mice injected with an irrelevant IgG at 24 hours post-infection were used as the control group.
    Figure Legend Snippet: Therapeutic efficacy of mAb 4E6 and 1H10 in a mouse model of H5N1 virus A/Vietnam/1194/04 infection. (A) The Kaplan-Meier survival curve. BALB/c mice (n=7 per group in two separate experiments) were infected intranasally with 10 LD 50 of A/Vietnam/1194/04 and at 24, 48, or 72 hours, were treated with a single intraperitoneal injection of mAb 4E6 or 1H10 at 10 mg/kg body weight. The control group received IgG at 10 mg/kg body weight at 24 hours post-infection. (B) Percentage weight change in mice treated with mAb 4E6 or 1H10 at different time points post-infection. The mice (n=7) were infected via intranasal inoculation with 10 LD50 of A/Vietnam/1203/4, followed by a single intraperitoneal injection of mAbs at 24, 48, or 72 hours post-infection, and their weights were monitored for 14 days. The mice injected with an irrelevant IgG at 24 hours post-infection were used as the control group.

    Techniques Used: Infection, Mouse Assay, Injection

    14) Product Images from "Dopamine-Dependent Tuning of Striatal Inhibitory Synaptogenesis"

    Article Title: Dopamine-Dependent Tuning of Striatal Inhibitory Synaptogenesis

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.4411-09.2010

    Dopamine receptor activation decreases the cell-surface expression of GABA A receptors via a dynamin-dependent pathway. A , B , Internalization of GABA A receptors was visualized by staining with β 2/3 -specific antibody in the presence of vehicle, SKF-38393 ( A ), or quinpirole ( B ), as described in Materials and Methods. Binding of β 2/3 -specific antibody to the surface expressed GABA A receptors was visualized using a secondary anti-mouse IgG coupled to Alexa555 (in red), while binding of the same antibody to internalized receptors was visualized using a secondary anti-mouse IgG coupled to Alexa488 (in green). Scale bars correspond to 10 μm. C , The decrease in surface β 2/3 subunit levels caused by SKF-38393 (SKF) was significantly attenuated in the presence of a dynamin inhibitory peptide (SKF/DynP), but not in the presence of its scrambled control (SKF/DynPC). D , The decrease in surface β 2/3 subunit levels caused by quinpirole (Q) was also significantly attenuated in the presence of a dynamin inhibitory peptide (Q/DynP), but not by its scrambled control (Q/DynPC). Bars represent mean ± SEM. Statistical analysis was performed using one-way ANOVA with Dunnett post hoc analysis versus control (* p
    Figure Legend Snippet: Dopamine receptor activation decreases the cell-surface expression of GABA A receptors via a dynamin-dependent pathway. A , B , Internalization of GABA A receptors was visualized by staining with β 2/3 -specific antibody in the presence of vehicle, SKF-38393 ( A ), or quinpirole ( B ), as described in Materials and Methods. Binding of β 2/3 -specific antibody to the surface expressed GABA A receptors was visualized using a secondary anti-mouse IgG coupled to Alexa555 (in red), while binding of the same antibody to internalized receptors was visualized using a secondary anti-mouse IgG coupled to Alexa488 (in green). Scale bars correspond to 10 μm. C , The decrease in surface β 2/3 subunit levels caused by SKF-38393 (SKF) was significantly attenuated in the presence of a dynamin inhibitory peptide (SKF/DynP), but not in the presence of its scrambled control (SKF/DynPC). D , The decrease in surface β 2/3 subunit levels caused by quinpirole (Q) was also significantly attenuated in the presence of a dynamin inhibitory peptide (Q/DynP), but not by its scrambled control (Q/DynPC). Bars represent mean ± SEM. Statistical analysis was performed using one-way ANOVA with Dunnett post hoc analysis versus control (* p

    Techniques Used: Activation Assay, Expressing, Staining, Binding Assay

    15) Product Images from "Integrin α2β1 Expression Regulates Matrix Metalloproteinase-1-Dependent Bronchial Epithelial Repair in Pulmonary Tuberculosis"

    Article Title: Integrin α2β1 Expression Regulates Matrix Metalloproteinase-1-Dependent Bronchial Epithelial Repair in Pulmonary Tuberculosis

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.01348

    Matrix type I collagen induces α2β1 integrin clustering and actin colocalization without affecting integrin surface expression. Human bronchial epithelial cells (HBECs) were cultured in the in the presence of matrix or soluble Coll-I or without extracellular matrix (ECM) as controls and stimulated with CoMtb or control medium. (A) Confocal microscopy figures of CoMtb-stimulated HBECs. Cells were stained with DAPI (blue) for nucleic acids, phalloidin conjugated with Alexa Fluor 594 (red) for F-actin, and FITC-conjugated anti-integrin α2 antibodies (green) for integrin subunit α2. Yellow color shows co-localization of F-actin and integrin α2β1 in merged images. Scale bar: 50 µm. (B,C) Control and CoMtb-stimulated HBECs were labeled with FITC-conjugated anti-integrin α2 or α3 and analyzed by FACS. FITC-conjugated mouse IgG1 was used as isotype control. Plots show mean fluorescence intensities (MFI) and SD for (B) α2 and (C) α3 integrins of three independent experiments.
    Figure Legend Snippet: Matrix type I collagen induces α2β1 integrin clustering and actin colocalization without affecting integrin surface expression. Human bronchial epithelial cells (HBECs) were cultured in the in the presence of matrix or soluble Coll-I or without extracellular matrix (ECM) as controls and stimulated with CoMtb or control medium. (A) Confocal microscopy figures of CoMtb-stimulated HBECs. Cells were stained with DAPI (blue) for nucleic acids, phalloidin conjugated with Alexa Fluor 594 (red) for F-actin, and FITC-conjugated anti-integrin α2 antibodies (green) for integrin subunit α2. Yellow color shows co-localization of F-actin and integrin α2β1 in merged images. Scale bar: 50 µm. (B,C) Control and CoMtb-stimulated HBECs were labeled with FITC-conjugated anti-integrin α2 or α3 and analyzed by FACS. FITC-conjugated mouse IgG1 was used as isotype control. Plots show mean fluorescence intensities (MFI) and SD for (B) α2 and (C) α3 integrins of three independent experiments.

    Techniques Used: Expressing, Cell Culture, Confocal Microscopy, Staining, Labeling, FACS, Fluorescence

    16) Product Images from "A Highly Sensitive Immunochromatographic Strip Test for Rapid and Quantitative Detection of Saikosaponin d"

    Article Title: A Highly Sensitive Immunochromatographic Strip Test for Rapid and Quantitative Detection of Saikosaponin d

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    doi: 10.3390/molecules23020338

    The structure and principle of the immunochromatographic strip (ICS). A nitrocellulose membrane, conjugate pad, sample pad, and absorbent pad were pasted onto a polyvinyl chloride backing to form the dipstick. Saikosaponin d (SSd)–bovine serum albumin (BSA) and goat anti-mouse IgG were used as the test capture reagent and control capture reagent, respectively. Using a dispenser, both the test and control capture reagents were dispensed separately as lines 0.5 cm apart on the nitrocellulose membrane.
    Figure Legend Snippet: The structure and principle of the immunochromatographic strip (ICS). A nitrocellulose membrane, conjugate pad, sample pad, and absorbent pad were pasted onto a polyvinyl chloride backing to form the dipstick. Saikosaponin d (SSd)–bovine serum albumin (BSA) and goat anti-mouse IgG were used as the test capture reagent and control capture reagent, respectively. Using a dispenser, both the test and control capture reagents were dispensed separately as lines 0.5 cm apart on the nitrocellulose membrane.

    Techniques Used: Stripping Membranes

    17) Product Images from "A Novel View on the Role of Intracellular Tails in Surface Delivery of the Potassium-Chloride Cotransporter KCC2"

    Article Title: A Novel View on the Role of Intracellular Tails in Surface Delivery of the Potassium-Chloride Cotransporter KCC2

    Journal: eNeuro

    doi: 10.1523/ENEURO.0055-17.2017

    Visualization of surface expressed and internalized KCC2-pH ext proteins using a live-cell immunolabeling protocol on cultured hippocampal neurons. A , Scheme of the multistep immunolabeling protocol applied to 13 DIV neurons. First Ab, primary antibody; second Cy3, Cy3-conjugated secondary antibody; second Alexa 647, Alexa Fluor 647–conjugated secondary antibody; PFA, paraformaldehyde. The scheme does not include final steps of fixed and permeabilized cells labeled with mouse anti-GFP and anti-mouse Alexa Fluor 488 antibody [total protein pool (F t )]. B , Representative images showing fluorescence emitted after staining with Cy3-conjugated secondary antibody [plasma membrane restricted pool (F m )]; images were pseudocolored using illustrated bicolor lookup table, first raw); Alexa Fluor 647–conjugated secondary antibody (internalized surface labeled molecules and portion of surface retained molecules, second raw); Alexa Fluor 488–conjugated secondary antibody (F t , third raw); internalized surface labeled signal obtained by arithmetic subtraction of first and second raw images (F i , fourth raw). Image columns illustrate fluorescent signals obtained at different z -planes or after arithmetic summation of nine planes as indicated. The neuronal shape (Alexa Fluor 488 fluorescence) is shown in light green in each image for reference. Insets illustrate indicated portions of images at higher zoom. Scale bars: 8 µm (main image), 1 µm (insets).
    Figure Legend Snippet: Visualization of surface expressed and internalized KCC2-pH ext proteins using a live-cell immunolabeling protocol on cultured hippocampal neurons. A , Scheme of the multistep immunolabeling protocol applied to 13 DIV neurons. First Ab, primary antibody; second Cy3, Cy3-conjugated secondary antibody; second Alexa 647, Alexa Fluor 647–conjugated secondary antibody; PFA, paraformaldehyde. The scheme does not include final steps of fixed and permeabilized cells labeled with mouse anti-GFP and anti-mouse Alexa Fluor 488 antibody [total protein pool (F t )]. B , Representative images showing fluorescence emitted after staining with Cy3-conjugated secondary antibody [plasma membrane restricted pool (F m )]; images were pseudocolored using illustrated bicolor lookup table, first raw); Alexa Fluor 647–conjugated secondary antibody (internalized surface labeled molecules and portion of surface retained molecules, second raw); Alexa Fluor 488–conjugated secondary antibody (F t , third raw); internalized surface labeled signal obtained by arithmetic subtraction of first and second raw images (F i , fourth raw). Image columns illustrate fluorescent signals obtained at different z -planes or after arithmetic summation of nine planes as indicated. The neuronal shape (Alexa Fluor 488 fluorescence) is shown in light green in each image for reference. Insets illustrate indicated portions of images at higher zoom. Scale bars: 8 µm (main image), 1 µm (insets).

    Techniques Used: Immunolabeling, Cell Culture, Labeling, Fluorescence, Staining

    18) Product Images from "A Highly Sensitive Immunochromatographic Strip Test for Rapid and Quantitative Detection of Saikosaponin d"

    Article Title: A Highly Sensitive Immunochromatographic Strip Test for Rapid and Quantitative Detection of Saikosaponin d

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    doi: 10.3390/molecules23020338

    The structure and principle of the immunochromatographic strip (ICS). A nitrocellulose membrane, conjugate pad, sample pad, and absorbent pad were pasted onto a polyvinyl chloride backing to form the dipstick. Saikosaponin d (SSd)–bovine serum albumin (BSA) and goat anti-mouse IgG were used as the test capture reagent and control capture reagent, respectively. Using a dispenser, both the test and control capture reagents were dispensed separately as lines 0.5 cm apart on the nitrocellulose membrane.
    Figure Legend Snippet: The structure and principle of the immunochromatographic strip (ICS). A nitrocellulose membrane, conjugate pad, sample pad, and absorbent pad were pasted onto a polyvinyl chloride backing to form the dipstick. Saikosaponin d (SSd)–bovine serum albumin (BSA) and goat anti-mouse IgG were used as the test capture reagent and control capture reagent, respectively. Using a dispenser, both the test and control capture reagents were dispensed separately as lines 0.5 cm apart on the nitrocellulose membrane.

    Techniques Used: Stripping Membranes

    19) Product Images from "Characterization of a Broadly Neutralizing Monoclonal Antibody That Targets the Fusion Domain of Group 2 Influenza A Virus Hemagglutinin"

    Article Title: Characterization of a Broadly Neutralizing Monoclonal Antibody That Targets the Fusion Domain of Group 2 Influenza A Virus Hemagglutinin

    Journal: Journal of Virology

    doi: 10.1128/JVI.02289-14

    MAb 9H10 is a pan-H3 HA antibody. ELISA plates were coated with 50 μl of purified baculovirus-expressed HA (2.5 μg/ml) and incubated overnight at 4°C. MAb 9H10 (starting at 100 μg/ml) was diluted 3-fold and incubated at room temperature for 2 h. The plates were washed thrice with 0.1% PBST, and a goat anti-mouse IgG antibody conjugated to HRP was used as a secondary antibody. Plates were incubated at 37°C for 1 h and then washed thrice with 0.1% PBST. The HRP substrate o -phenylenediamine dihydrochloride was used, and plates were read at 450 nm. A control IgG (an anti-ankyrin MAb, ANK) was used as a negative control, and MAb 12D1, a previously identified pan-H3 MAb, was used as a positive control. A nonlinear curved was generated with GraphPad Prism 4.0.
    Figure Legend Snippet: MAb 9H10 is a pan-H3 HA antibody. ELISA plates were coated with 50 μl of purified baculovirus-expressed HA (2.5 μg/ml) and incubated overnight at 4°C. MAb 9H10 (starting at 100 μg/ml) was diluted 3-fold and incubated at room temperature for 2 h. The plates were washed thrice with 0.1% PBST, and a goat anti-mouse IgG antibody conjugated to HRP was used as a secondary antibody. Plates were incubated at 37°C for 1 h and then washed thrice with 0.1% PBST. The HRP substrate o -phenylenediamine dihydrochloride was used, and plates were read at 450 nm. A control IgG (an anti-ankyrin MAb, ANK) was used as a negative control, and MAb 12D1, a previously identified pan-H3 MAb, was used as a positive control. A nonlinear curved was generated with GraphPad Prism 4.0.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Purification, Incubation, Negative Control, Positive Control, Generated

    20) Product Images from "Tomato HsfA1a plays a critical role in plant drought tolerance by activating ATG genes and inducing autophagy"

    Article Title: Tomato HsfA1a plays a critical role in plant drought tolerance by activating ATG genes and inducing autophagy

    Journal: Autophagy

    doi: 10.1080/15548627.2015.1098798

    HsfA1a binds to the promoters of ATG10 and ATG18f in vitro and in vivo . ( A ) Oligonucleotide used in the electrophoretic mobility shift assays (EMSA). The ATG10 probe contains one direct HSE, whereas in the ATG10−1Δ and ATG10–2Δ probes the HSE core sequence was mutated. The ATG18f probe contains one direct HSE, whereas in the ATG18fΔ probe, the HSE core sequence was mutated. The WT and mutated HSE sequences are underlined. The mutated bases were indicated in red. ( B and C ) EMSA showing HsfA1a bound to the HSE sequences of the ATG10 or ATG18f promoters. Recombinant HsfA1a was purified from E. coli cells and used for DNA binding assays with ATG10, ATG10−1Δ, ATG10–2Δ, ATG18f, or ATG18fΔ as the probes. His was included as the negative control. ( D ) Direct binding of HsfA1a to the ATG10 and ATG18f promoters was analyzed using ChIP-qPCR in 35S-HsfA1a-HA -overexpressing ( HsfA1a OE) plants. Six-wk-old HsfA1a OE plants were exposed to dehydration by withholding water and input chromatin was isolated from leaf samples on d 6. The epitope-tagged HsfA1a-chromatin complex was immunoprecipitated with an anti-HA antibody. A control reaction was processed side-by-side using mouse IgG. Input- and ChIP-DNA samples were quantified by qRT-PCR using primers specific for the promoters of the ATG genes. The ChIP results are presented as percentage of the input DNA. Means with the same letter did not significantly differ at P
    Figure Legend Snippet: HsfA1a binds to the promoters of ATG10 and ATG18f in vitro and in vivo . ( A ) Oligonucleotide used in the electrophoretic mobility shift assays (EMSA). The ATG10 probe contains one direct HSE, whereas in the ATG10−1Δ and ATG10–2Δ probes the HSE core sequence was mutated. The ATG18f probe contains one direct HSE, whereas in the ATG18fΔ probe, the HSE core sequence was mutated. The WT and mutated HSE sequences are underlined. The mutated bases were indicated in red. ( B and C ) EMSA showing HsfA1a bound to the HSE sequences of the ATG10 or ATG18f promoters. Recombinant HsfA1a was purified from E. coli cells and used for DNA binding assays with ATG10, ATG10−1Δ, ATG10–2Δ, ATG18f, or ATG18fΔ as the probes. His was included as the negative control. ( D ) Direct binding of HsfA1a to the ATG10 and ATG18f promoters was analyzed using ChIP-qPCR in 35S-HsfA1a-HA -overexpressing ( HsfA1a OE) plants. Six-wk-old HsfA1a OE plants were exposed to dehydration by withholding water and input chromatin was isolated from leaf samples on d 6. The epitope-tagged HsfA1a-chromatin complex was immunoprecipitated with an anti-HA antibody. A control reaction was processed side-by-side using mouse IgG. Input- and ChIP-DNA samples were quantified by qRT-PCR using primers specific for the promoters of the ATG genes. The ChIP results are presented as percentage of the input DNA. Means with the same letter did not significantly differ at P

    Techniques Used: In Vitro, In Vivo, Electrophoretic Mobility Shift Assay, Sequencing, Recombinant, Purification, Binding Assay, Negative Control, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Isolation, Immunoprecipitation, Quantitative RT-PCR

    21) Product Images from "Exogenous Secreted Frizzled-Related Protein-4 Modulates Steroidogenesis of Rat Granulosa Cells through Wnt/β-catenin and PI3K/AKT Signaling Pathways"

    Article Title: Exogenous Secreted Frizzled-Related Protein-4 Modulates Steroidogenesis of Rat Granulosa Cells through Wnt/β-catenin and PI3K/AKT Signaling Pathways

    Journal: Avicenna Journal of Medical Biotechnology

    doi:

    Active β-catenin and cleaved caspase-3 are co-localized in FSH-primed but not LH-stimulated granulosa cells (GCs). Cells were double immunostained with anti-active β-catenin (red) and anti-active casapse-3 (green). DNA was observed by using DAPI (blue). A–C) Cytoplasmic immunostaining of active β-catenin and active caspase-3 in untreated (control) GCs. D–F) Nuclear localization of active β-catenin and active caspase-3 in FSH-primed GCs. G–I) Cytoplasmic immunostaining of active β-catenin and active caspase-3 in LH-stimulated cells. J–K) The negative control was performed by using IgG1 mouse isotypic control as primary antibody. Images are at 200× magnification.
    Figure Legend Snippet: Active β-catenin and cleaved caspase-3 are co-localized in FSH-primed but not LH-stimulated granulosa cells (GCs). Cells were double immunostained with anti-active β-catenin (red) and anti-active casapse-3 (green). DNA was observed by using DAPI (blue). A–C) Cytoplasmic immunostaining of active β-catenin and active caspase-3 in untreated (control) GCs. D–F) Nuclear localization of active β-catenin and active caspase-3 in FSH-primed GCs. G–I) Cytoplasmic immunostaining of active β-catenin and active caspase-3 in LH-stimulated cells. J–K) The negative control was performed by using IgG1 mouse isotypic control as primary antibody. Images are at 200× magnification.

    Techniques Used: Immunostaining, Negative Control

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    Autoradiography:

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    Enzyme-linked Immunosorbent Assay:

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    Article Snippet: .. C5-1 Quantification Enzyme-linked immunosorbent assay (ELISA) plates for C5-1 quantification (Becton Dickinson, Mississauga ON, Canada) were coated with 3.75 µg/mL goat anti-mouse heavy chain-specific IgG1 (Sigma-Aldrich) in 50 mM carbonate buffer (pH 9.0) at 4°C for 16–18 h. The plates were washed three times in 10 mM phosphate-buffered saline containing 0.1% (v/v) Tween 20 (PBS-T), blocked through a 1-h incubation in 1% (w/v) casein in phosphate-buffered saline (PBS) (Pierce, Rockford IL, USA) at 37°C, and washed three times in PBS-T. A standard curve was generated for each plate with 0, 4, 8, 16, 24, 32, 40 and 60 ng/mL of purified mouse IgG1 (Sigma-Aldrich). ..

    Article Title: SHCBP1 regulates apoptosis in lung cancer cells through phosphatase and tensin homolog
    Article Snippet: .. Following washing, the membrane was incubated with secondary anti-rabbit immunoglobulin G (IgG; 1:1,000; cat. no. A3687; Sigma-Aldrich; Merck KGaA) and anti-mouse IgG (1:1,000; cat. no. M8770; Sigma-Aldrich; Merck KGaA) antibodies at room temperature for 1 h. Images were captured on an Odyssey CLx Infrared Imaging system (LI-COR Biosciences). .. Subsequently, the blots were semi-quantified using Odyssey CLx 2.1 software (LI-COR Biosciences).

    Article Title: Capsular Polysaccharide-Fimbrial Protein Conjugate Vaccine Protects against Porphyromonas gingivalis Infection in SCID Mice Reconstituted with Human Peripheral Blood Lymphocytes
    Article Snippet: .. After being washed three times with PBS containing 0.1% Tween 20, 0.05 ml of goat anti-mouse IgG (heavy- and light-chain specific, affinity purified, alkaline phosphatase conjugated; Calbiochem, Basel, Switzerland) diluted in PBS containing 0.1% Tween 20 were added to each well and incubated overnight at room temperature. .. After the plates were washed, 0.1 ml of nitrophenyl phosphate (1 mg/ml in diethanolamine buffer, pH 9.8) was added to each well and incubated for 60 min, and 0.1 ml of 3 N NaOH was added to stop the color reaction.

    Article Title: A small heat shock/?-crystallin protein from encysted Artemia embryos suppresses tubulin denaturation
    Article Snippet: .. Membranes were washed and then incubated for 15 minutes at room temperature, with peroxidase-conjugated secondary antibodies diluted in 10 mM Tris-HCl, 1 M NaCl, 0.5% (v/v) Tween 20, pH 7.4 (HST), including goat anti-rabbit IgG (Bio/Can Scientific, Mississauga, Ontario, Canada), goat anti-mouse IgG (Sigma) and mouse anti-rabbit IgG, and γ-chain–specific clone RG-96 (Sigma), which recognizes only the heavy chain of IgG. .. Blots were washed after secondary antibody exposure, and immunoreactive proteins were revealed by use of the enhanced chemiluminescence technique (Renaissance®, NEM™, Life Science Products Inc, Boston, MA, USA) and exposure to autoradiography film (Labscientific, Livingston, NJ, USA).

    Article Title: Extensive Post-translational Modification of Active and Inactivated Forms of Endogenous p53 *
    Article Snippet: Immunoprecipitation efficiency and p53 protein concentration were assessed via immunoblotting as described previously ( ). p53 was visualized with mouse monoclonal antibody D01-HRP (sc-126, Santa Cruz Biotechnologies, Santa Cruz, CA), actin with mouse monoclonal antibody AC-15 HRP (Abcam, Cambridge, MA), and IgG heavy chain with goat anti-mouse HRP (AP124P, Chemicon, EMD Millipore, Billerica, MA). .. Cells were maintained in the presence of etoposide for 44 h, washed twice in PBS, harvested, and incubated in lysis buffer containing N -lauryl sarcosine for analysis via immunoblotting.

    Article Title: Tracking the Emerging Human Pathogen Pseudallescheria boydii by Using Highly Specific Monoclonal Antibodies ▿
    Article Snippet: .. Antigen extracts prepared in PBST as described above were incubated in the antibody-coated wells for 2 h. The wells were given four 5-min rinses with PBST and then incubated sequentially with HG12 MAb (IgG1) supernatant for 1 h, followed by goat anti-mouse IgG (γ-chain specific) peroxidase conjugate (Sigma), diluted 1 in 1,000 in PBST, for a further hour. .. Working volumes were 50 μl per well, and control wells were incubated with PBST antigen extract from P. boydii grown on semiselective medium under the same conditions.

    BIA-KA:

    Article Title: SHCBP1 regulates apoptosis in lung cancer cells through phosphatase and tensin homolog
    Article Snippet: The concentration of the proteins was determined using a BCA Protein Assay kit (Beyotime Institute of Biotechnology). .. Following washing, the membrane was incubated with secondary anti-rabbit immunoglobulin G (IgG; 1:1,000; cat. no. A3687; Sigma-Aldrich; Merck KGaA) and anti-mouse IgG (1:1,000; cat. no. M8770; Sigma-Aldrich; Merck KGaA) antibodies at room temperature for 1 h. Images were captured on an Odyssey CLx Infrared Imaging system (LI-COR Biosciences).

    Modification:

    Article Title: Simvastatin Efficiently Lowers Small LDL-IgG Immune Complex Levels: A Therapeutic Quality beyond the Lipid-Lowering Effect
    Article Snippet: Paragraph title: Supporting Information DELFIA setup and analysis. Iodixanol gradient ultracentrifugation of human plasma and free proteins. Agarose electrophoresis of lipoprotein fractions isolated after iodixanol gradient ultracentrifugation and immunodetection of human IgG. Detection of oxidatively modified LDL with the monoclonal antibody OB/04 (directed against oxidation-specific epitopes). Distribution of small LDL-IgG-IC in LDL-subfractions isolated from 11 CAD patients. ... Fluorescence counts (1 μg OB/04 antibody per well) were recorded after incubation with europium labelled goat anti-mouse IgG (#M-8770, Sigma Immunochemicals, St. Louis; USA).

    Western Blot:

    Article Title: SHCBP1 regulates apoptosis in lung cancer cells through phosphatase and tensin homolog
    Article Snippet: Paragraph title: Western blot analysis ... Following washing, the membrane was incubated with secondary anti-rabbit immunoglobulin G (IgG; 1:1,000; cat. no. A3687; Sigma-Aldrich; Merck KGaA) and anti-mouse IgG (1:1,000; cat. no. M8770; Sigma-Aldrich; Merck KGaA) antibodies at room temperature for 1 h. Images were captured on an Odyssey CLx Infrared Imaging system (LI-COR Biosciences).

    Article Title: A small heat shock/?-crystallin protein from encysted Artemia embryos suppresses tubulin denaturation
    Article Snippet: Paragraph title: Electrophoresis, Western blotting, and protein immunodetection ... Membranes were washed and then incubated for 15 minutes at room temperature, with peroxidase-conjugated secondary antibodies diluted in 10 mM Tris-HCl, 1 M NaCl, 0.5% (v/v) Tween 20, pH 7.4 (HST), including goat anti-rabbit IgG (Bio/Can Scientific, Mississauga, Ontario, Canada), goat anti-mouse IgG (Sigma) and mouse anti-rabbit IgG, and γ-chain–specific clone RG-96 (Sigma), which recognizes only the heavy chain of IgG.

    Flow Cytometry:

    Article Title: Tracking the Emerging Human Pathogen Pseudallescheria boydii by Using Highly Specific Monoclonal Antibodies ▿
    Article Snippet: Wells were washed three times (5 min each time) with PBST, washed once with PBS and once with dH2 O, and air dried at 23°C with a laminar flow hood. .. Antigen extracts prepared in PBST as described above were incubated in the antibody-coated wells for 2 h. The wells were given four 5-min rinses with PBST and then incubated sequentially with HG12 MAb (IgG1) supernatant for 1 h, followed by goat anti-mouse IgG (γ-chain specific) peroxidase conjugate (Sigma), diluted 1 in 1,000 in PBST, for a further hour.

    Concentration Assay:

    Article Title: Simvastatin Efficiently Lowers Small LDL-IgG Immune Complex Levels: A Therapeutic Quality beyond the Lipid-Lowering Effect
    Article Snippet: Fluorescence counts (1 μg OB/04 antibody per well) were recorded after incubation with europium labelled goat anti-mouse IgG (#M-8770, Sigma Immunochemicals, St. Louis; USA). .. OB/04 DELFIA counts divided by the corresponding fluorescence counts of the apoB represent the concentration of modified apoB normalized to the concentration of apoB.

    Article Title: SHCBP1 regulates apoptosis in lung cancer cells through phosphatase and tensin homolog
    Article Snippet: The concentration of the proteins was determined using a BCA Protein Assay kit (Beyotime Institute of Biotechnology). .. Following washing, the membrane was incubated with secondary anti-rabbit immunoglobulin G (IgG; 1:1,000; cat. no. A3687; Sigma-Aldrich; Merck KGaA) and anti-mouse IgG (1:1,000; cat. no. M8770; Sigma-Aldrich; Merck KGaA) antibodies at room temperature for 1 h. Images were captured on an Odyssey CLx Infrared Imaging system (LI-COR Biosciences).

    Article Title: Extensive Post-translational Modification of Active and Inactivated Forms of Endogenous p53 *
    Article Snippet: Immunoprecipitation efficiency and p53 protein concentration were assessed via immunoblotting as described previously ( ). p53 was visualized with mouse monoclonal antibody D01-HRP (sc-126, Santa Cruz Biotechnologies, Santa Cruz, CA), actin with mouse monoclonal antibody AC-15 HRP (Abcam, Cambridge, MA), and IgG heavy chain with goat anti-mouse HRP (AP124P, Chemicon, EMD Millipore, Billerica, MA). .. For the comparison of p53 protein concentrations in infected and etoposide-treated cells, etoposide (Sigma) was diluted to 50 m m in dimethyl sulfoxide and added to medium at a final concentration of 125 μ m .

    Protease Inhibitor:

    Article Title: SHCBP1 regulates apoptosis in lung cancer cells through phosphatase and tensin homolog
    Article Snippet: Protein samples (70 µg) were extracted with lysis buffer (Beyotime Institute of Biotechnology) supplemented with 1% protease inhibitor solution. .. Following washing, the membrane was incubated with secondary anti-rabbit immunoglobulin G (IgG; 1:1,000; cat. no. A3687; Sigma-Aldrich; Merck KGaA) and anti-mouse IgG (1:1,000; cat. no. M8770; Sigma-Aldrich; Merck KGaA) antibodies at room temperature for 1 h. Images were captured on an Odyssey CLx Infrared Imaging system (LI-COR Biosciences).

    Infection:

    Article Title: Capsular Polysaccharide-Fimbrial Protein Conjugate Vaccine Protects against Porphyromonas gingivalis Infection in SCID Mice Reconstituted with Human Peripheral Blood Lymphocytes
    Article Snippet: Preimmune, postimmune (2 weeks following final immunization), and postinfection (3 weeks following infection) total IgG and IgG subclass antibody titers were determined by enzyme-linked immunosorbent assay (ELISA) with an alkaline phosphatase assay system. .. After being washed three times with PBS containing 0.1% Tween 20, 0.05 ml of goat anti-mouse IgG (heavy- and light-chain specific, affinity purified, alkaline phosphatase conjugated; Calbiochem, Basel, Switzerland) diluted in PBS containing 0.1% Tween 20 were added to each well and incubated overnight at room temperature.

    Article Title: Extensive Post-translational Modification of Active and Inactivated Forms of Endogenous p53 *
    Article Snippet: Immunoprecipitation efficiency and p53 protein concentration were assessed via immunoblotting as described previously ( ). p53 was visualized with mouse monoclonal antibody D01-HRP (sc-126, Santa Cruz Biotechnologies, Santa Cruz, CA), actin with mouse monoclonal antibody AC-15 HRP (Abcam, Cambridge, MA), and IgG heavy chain with goat anti-mouse HRP (AP124P, Chemicon, EMD Millipore, Billerica, MA). .. For the comparison of p53 protein concentrations in infected and etoposide-treated cells, etoposide (Sigma) was diluted to 50 m m in dimethyl sulfoxide and added to medium at a final concentration of 125 μ m .

    Generated:

    Article Title: Protection of Recombinant Mammalian Antibodies from Development-Dependent Proteolysis in Leaves of Nicotiana benthamiana
    Article Snippet: .. C5-1 Quantification Enzyme-linked immunosorbent assay (ELISA) plates for C5-1 quantification (Becton Dickinson, Mississauga ON, Canada) were coated with 3.75 µg/mL goat anti-mouse heavy chain-specific IgG1 (Sigma-Aldrich) in 50 mM carbonate buffer (pH 9.0) at 4°C for 16–18 h. The plates were washed three times in 10 mM phosphate-buffered saline containing 0.1% (v/v) Tween 20 (PBS-T), blocked through a 1-h incubation in 1% (w/v) casein in phosphate-buffered saline (PBS) (Pierce, Rockford IL, USA) at 37°C, and washed three times in PBS-T. A standard curve was generated for each plate with 0, 4, 8, 16, 24, 32, 40 and 60 ng/mL of purified mouse IgG1 (Sigma-Aldrich). ..

    other:

    Article Title: Expression of glioma-associated antigens in pediatric brain stem and non-brain stem gliomas
    Article Snippet: The secondary antibodies used in the current study were peroxidase-conjugated rabbit polyclonal anti-goat IgG heavy and light chains (1:200; ab6741, Abcam), goat polyclonal anti-mouse IgG heavy and light chains (1:200; AP124P, Upstate Cell Signaling) and goat polyclonal anti-rabbit IgG heavy and light chains (1:500; AP132P, Chemicon International).

    Article Title: A conserved quality-control pathway that mediates degradation of unassembled ribosomal proteins
    Article Snippet: For secondary antibody, HRP-conjugated anti-rabbit IgG (A6154; RRID: AB_258284 ; 1:10,000 dilution) and HRP-conjugated anti-mouse IgG (M8770; RRID: AB_260711 ; 1:10,000 dilution) were from Sigma, IR680RD conjugated anti-rabbit (926–68071; RRID: AB_10956166 ; 1:10,000 dilution) and IR800CW conjugated anti-mouse (926–32210; RRID: AB_621842 ; 1:10,000 dilution) were from LI-COR Biosciences (Lincoln, NE).

    Article Title: Use of an Attenuated Leishmanial Parasite as an Immunoprophylactic and Immunotherapeutic Agent against Murine Visceral Leishmaniasis
    Article Snippet: Mouse IgG1 and goat anti-mouse IgG1 (heavy chain specific) were purchased from Sigma, St. Louis, Mo.

    Article Title: Ribosomal proteins produced in excess are degraded by the ubiquitin–proteasome system
    Article Snippet: For secondary antibody, horseradish peroxidase (HRP)–conjugated anti-rabbit immunoglobulin G (IgG; A6154; 1:10,000 dilution) and HRP-conjugated anti-mouse IgG (M8770; 1:10,000 dilution) were from Sigma-Aldrich, and IR680RD-conjugated anti-rabbit (926-68071; 1:10,000 dilution) and IR800CW-conjugated anti-mouse (926-32210; 1:10,000 dilution) were from LI-COR Biotechnology (Lincoln, NE).

    Imaging:

    Article Title: SHCBP1 regulates apoptosis in lung cancer cells through phosphatase and tensin homolog
    Article Snippet: .. Following washing, the membrane was incubated with secondary anti-rabbit immunoglobulin G (IgG; 1:1,000; cat. no. A3687; Sigma-Aldrich; Merck KGaA) and anti-mouse IgG (1:1,000; cat. no. M8770; Sigma-Aldrich; Merck KGaA) antibodies at room temperature for 1 h. Images were captured on an Odyssey CLx Infrared Imaging system (LI-COR Biosciences). .. Subsequently, the blots were semi-quantified using Odyssey CLx 2.1 software (LI-COR Biosciences).

    Protein Concentration:

    Article Title: Extensive Post-translational Modification of Active and Inactivated Forms of Endogenous p53 *
    Article Snippet: .. Immunoprecipitation efficiency and p53 protein concentration were assessed via immunoblotting as described previously ( ). p53 was visualized with mouse monoclonal antibody D01-HRP (sc-126, Santa Cruz Biotechnologies, Santa Cruz, CA), actin with mouse monoclonal antibody AC-15 HRP (Abcam, Cambridge, MA), and IgG heavy chain with goat anti-mouse HRP (AP124P, Chemicon, EMD Millipore, Billerica, MA). ..

    Affinity Purification:

    Article Title: Simvastatin Efficiently Lowers Small LDL-IgG Immune Complex Levels: A Therapeutic Quality beyond the Lipid-Lowering Effect
    Article Snippet: Affinity purified small LDL-IgG-IC (bound) and residual LDL (unbound; small LDL-IgG-IC depleted) were prepared from an LDL-subfraction rich in small LDL-IgG-IC. .. Fluorescence counts (1 μg OB/04 antibody per well) were recorded after incubation with europium labelled goat anti-mouse IgG (#M-8770, Sigma Immunochemicals, St. Louis; USA).

    Article Title: Capsular Polysaccharide-Fimbrial Protein Conjugate Vaccine Protects against Porphyromonas gingivalis Infection in SCID Mice Reconstituted with Human Peripheral Blood Lymphocytes
    Article Snippet: .. After being washed three times with PBS containing 0.1% Tween 20, 0.05 ml of goat anti-mouse IgG (heavy- and light-chain specific, affinity purified, alkaline phosphatase conjugated; Calbiochem, Basel, Switzerland) diluted in PBS containing 0.1% Tween 20 were added to each well and incubated overnight at room temperature. .. After the plates were washed, 0.1 ml of nitrophenyl phosphate (1 mg/ml in diethanolamine buffer, pH 9.8) was added to each well and incubated for 60 min, and 0.1 ml of 3 N NaOH was added to stop the color reaction.

    Immunofluorescence:

    Article Title: Exendin-4 Ameliorates Lipotoxicity-induced Glomerular Endothelial Cell Injury by Improving ABC Transporter A1-mediated Cholesterol Efflux in Diabetic apoE Knockout Mice *
    Article Snippet: Paragraph title: Immunofluorescence ... The sections were then stained with FITC-labeled rabbit anti-goat immunoglobulin (Dako), Tetramethylrhodamine isothiocyanate-labeled rabbit anti-mouse immunoglobulin (Dako), FITC-stained swine anti-rabbit immunoglobulin (Dako), or goat anti-mouse IgG (heavy and light chain) (Millipore).

    Molecular Weight:

    Article Title: A small heat shock/?-crystallin protein from encysted Artemia embryos suppresses tubulin denaturation
    Article Snippet: Molecular weight standards were from BioRad Laboratories Ltd. Proteins were also transferred to Protram™ nitrocellulose membranes (Mandel Scientific Co. Ltd) after electrophoresis ( ). .. Membranes were washed and then incubated for 15 minutes at room temperature, with peroxidase-conjugated secondary antibodies diluted in 10 mM Tris-HCl, 1 M NaCl, 0.5% (v/v) Tween 20, pH 7.4 (HST), including goat anti-rabbit IgG (Bio/Can Scientific, Mississauga, Ontario, Canada), goat anti-mouse IgG (Sigma) and mouse anti-rabbit IgG, and γ-chain–specific clone RG-96 (Sigma), which recognizes only the heavy chain of IgG.

    Fluorescence:

    Article Title: Simvastatin Efficiently Lowers Small LDL-IgG Immune Complex Levels: A Therapeutic Quality beyond the Lipid-Lowering Effect
    Article Snippet: .. Fluorescence counts (1 μg OB/04 antibody per well) were recorded after incubation with europium labelled goat anti-mouse IgG (#M-8770, Sigma Immunochemicals, St. Louis; USA). .. OB/04 DELFIA counts divided by the corresponding fluorescence counts of the apoB represent the concentration of modified apoB normalized to the concentration of apoB.

    Isolation:

    Article Title: Simvastatin Efficiently Lowers Small LDL-IgG Immune Complex Levels: A Therapeutic Quality beyond the Lipid-Lowering Effect
    Article Snippet: Paragraph title: Supporting Information DELFIA setup and analysis. Iodixanol gradient ultracentrifugation of human plasma and free proteins. Agarose electrophoresis of lipoprotein fractions isolated after iodixanol gradient ultracentrifugation and immunodetection of human IgG. Detection of oxidatively modified LDL with the monoclonal antibody OB/04 (directed against oxidation-specific epitopes). Distribution of small LDL-IgG-IC in LDL-subfractions isolated from 11 CAD patients. ... Fluorescence counts (1 μg OB/04 antibody per well) were recorded after incubation with europium labelled goat anti-mouse IgG (#M-8770, Sigma Immunochemicals, St. Louis; USA).

    Immunodetection:

    Article Title: Simvastatin Efficiently Lowers Small LDL-IgG Immune Complex Levels: A Therapeutic Quality beyond the Lipid-Lowering Effect
    Article Snippet: Paragraph title: Supporting Information DELFIA setup and analysis. Iodixanol gradient ultracentrifugation of human plasma and free proteins. Agarose electrophoresis of lipoprotein fractions isolated after iodixanol gradient ultracentrifugation and immunodetection of human IgG. Detection of oxidatively modified LDL with the monoclonal antibody OB/04 (directed against oxidation-specific epitopes). Distribution of small LDL-IgG-IC in LDL-subfractions isolated from 11 CAD patients. ... Fluorescence counts (1 μg OB/04 antibody per well) were recorded after incubation with europium labelled goat anti-mouse IgG (#M-8770, Sigma Immunochemicals, St. Louis; USA).

    Article Title: A small heat shock/?-crystallin protein from encysted Artemia embryos suppresses tubulin denaturation
    Article Snippet: Paragraph title: Electrophoresis, Western blotting, and protein immunodetection ... Membranes were washed and then incubated for 15 minutes at room temperature, with peroxidase-conjugated secondary antibodies diluted in 10 mM Tris-HCl, 1 M NaCl, 0.5% (v/v) Tween 20, pH 7.4 (HST), including goat anti-rabbit IgG (Bio/Can Scientific, Mississauga, Ontario, Canada), goat anti-mouse IgG (Sigma) and mouse anti-rabbit IgG, and γ-chain–specific clone RG-96 (Sigma), which recognizes only the heavy chain of IgG.

    Purification:

    Article Title: Protection of Recombinant Mammalian Antibodies from Development-Dependent Proteolysis in Leaves of Nicotiana benthamiana
    Article Snippet: .. C5-1 Quantification Enzyme-linked immunosorbent assay (ELISA) plates for C5-1 quantification (Becton Dickinson, Mississauga ON, Canada) were coated with 3.75 µg/mL goat anti-mouse heavy chain-specific IgG1 (Sigma-Aldrich) in 50 mM carbonate buffer (pH 9.0) at 4°C for 16–18 h. The plates were washed three times in 10 mM phosphate-buffered saline containing 0.1% (v/v) Tween 20 (PBS-T), blocked through a 1-h incubation in 1% (w/v) casein in phosphate-buffered saline (PBS) (Pierce, Rockford IL, USA) at 37°C, and washed three times in PBS-T. A standard curve was generated for each plate with 0, 4, 8, 16, 24, 32, 40 and 60 ng/mL of purified mouse IgG1 (Sigma-Aldrich). ..

    Article Title: Extensive Post-translational Modification of Active and Inactivated Forms of Endogenous p53 *
    Article Snippet: Paragraph title: p53 Purification and Immunoprecipitation ... Immunoprecipitation efficiency and p53 protein concentration were assessed via immunoblotting as described previously ( ). p53 was visualized with mouse monoclonal antibody D01-HRP (sc-126, Santa Cruz Biotechnologies, Santa Cruz, CA), actin with mouse monoclonal antibody AC-15 HRP (Abcam, Cambridge, MA), and IgG heavy chain with goat anti-mouse HRP (AP124P, Chemicon, EMD Millipore, Billerica, MA).

    ALP Assay:

    Article Title: Capsular Polysaccharide-Fimbrial Protein Conjugate Vaccine Protects against Porphyromonas gingivalis Infection in SCID Mice Reconstituted with Human Peripheral Blood Lymphocytes
    Article Snippet: Preimmune, postimmune (2 weeks following final immunization), and postinfection (3 weeks following infection) total IgG and IgG subclass antibody titers were determined by enzyme-linked immunosorbent assay (ELISA) with an alkaline phosphatase assay system. .. After being washed three times with PBS containing 0.1% Tween 20, 0.05 ml of goat anti-mouse IgG (heavy- and light-chain specific, affinity purified, alkaline phosphatase conjugated; Calbiochem, Basel, Switzerland) diluted in PBS containing 0.1% Tween 20 were added to each well and incubated overnight at room temperature.

    Staining:

    Article Title: A small heat shock/?-crystallin protein from encysted Artemia embryos suppresses tubulin denaturation
    Article Snippet: Blots were stained with Ponceau S (Sigma) to ensure efficient protein transfer, destained, and probed. .. Membranes were washed and then incubated for 15 minutes at room temperature, with peroxidase-conjugated secondary antibodies diluted in 10 mM Tris-HCl, 1 M NaCl, 0.5% (v/v) Tween 20, pH 7.4 (HST), including goat anti-rabbit IgG (Bio/Can Scientific, Mississauga, Ontario, Canada), goat anti-mouse IgG (Sigma) and mouse anti-rabbit IgG, and γ-chain–specific clone RG-96 (Sigma), which recognizes only the heavy chain of IgG.

    Article Title: Exendin-4 Ameliorates Lipotoxicity-induced Glomerular Endothelial Cell Injury by Improving ABC Transporter A1-mediated Cholesterol Efflux in Diabetic apoE Knockout Mice *
    Article Snippet: .. The sections were then stained with FITC-labeled rabbit anti-goat immunoglobulin (Dako), Tetramethylrhodamine isothiocyanate-labeled rabbit anti-mouse immunoglobulin (Dako), FITC-stained swine anti-rabbit immunoglobulin (Dako), or goat anti-mouse IgG (heavy and light chain) (Millipore). .. The tissue sections were analyzed with a Nikon A1Si confocal laser scanning microscope and NIS-Elements Advanced Research software, version 4.0 (Nikon, Tokyo), and S Plan Fluor ELWD 40× DIC.

    Software:

    Article Title: SHCBP1 regulates apoptosis in lung cancer cells through phosphatase and tensin homolog
    Article Snippet: Following washing, the membrane was incubated with secondary anti-rabbit immunoglobulin G (IgG; 1:1,000; cat. no. A3687; Sigma-Aldrich; Merck KGaA) and anti-mouse IgG (1:1,000; cat. no. M8770; Sigma-Aldrich; Merck KGaA) antibodies at room temperature for 1 h. Images were captured on an Odyssey CLx Infrared Imaging system (LI-COR Biosciences). .. Subsequently, the blots were semi-quantified using Odyssey CLx 2.1 software (LI-COR Biosciences).

    Article Title: Exendin-4 Ameliorates Lipotoxicity-induced Glomerular Endothelial Cell Injury by Improving ABC Transporter A1-mediated Cholesterol Efflux in Diabetic apoE Knockout Mice *
    Article Snippet: The sections were then stained with FITC-labeled rabbit anti-goat immunoglobulin (Dako), Tetramethylrhodamine isothiocyanate-labeled rabbit anti-mouse immunoglobulin (Dako), FITC-stained swine anti-rabbit immunoglobulin (Dako), or goat anti-mouse IgG (heavy and light chain) (Millipore). .. The tissue sections were analyzed with a Nikon A1Si confocal laser scanning microscope and NIS-Elements Advanced Research software, version 4.0 (Nikon, Tokyo), and S Plan Fluor ELWD 40× DIC.

    ROS Assay:

    Article Title: Melatonin receptor stimulation by agomelatine prevents Aβ-induced tau phosphorylation and oxidative damage in PC12 cells
    Article Snippet: Aβ25–35 (#A4559), agomelatine (#A1362), luzindole (#L2407), the primary antibodies against phosphotau (Ser396) (#SAB4504557), tau (#SAB4501830), PTEN (#SAB1406331), GAPDH (#SAB2701826), goat antirab-bit IgG (#A3687), and antibody antimouse IgG (#M8770) were purchased from Sigma-Aldrich Co., St Louis, MO, USA. .. Cell counting kit-8 (CCK-8) (#E606335-0500) and ROS assay kit (#50101ES01) were obtained from Sango Biotech (Shanghai, China).

    Article Title: Melatonin receptor stimulation by agomelatine prevents Aβ-induced tau phosphorylation and oxidative damage in PC12 cells
    Article Snippet: Materials Aβ25–35 (#A4559), agomelatine (#A1362), luzindole (#L2407), the primary antibodies against phosphotau (Ser396) (#SAB4504557), tau (#SAB4501830), PTEN (#SAB1406331), GAPDH (#SAB2701826), goat antirab-bit IgG (#A3687), and antibody antimouse IgG (#M8770) were purchased from Sigma-Aldrich Co., St Louis, MO, USA. .. Cell counting kit-8 (CCK-8) (#E606335-0500) and ROS assay kit (#50101ES01) were obtained from Sango Biotech (Shanghai, China).

    Electrophoresis:

    Article Title: Simvastatin Efficiently Lowers Small LDL-IgG Immune Complex Levels: A Therapeutic Quality beyond the Lipid-Lowering Effect
    Article Snippet: Paragraph title: Supporting Information DELFIA setup and analysis. Iodixanol gradient ultracentrifugation of human plasma and free proteins. Agarose electrophoresis of lipoprotein fractions isolated after iodixanol gradient ultracentrifugation and immunodetection of human IgG. Detection of oxidatively modified LDL with the monoclonal antibody OB/04 (directed against oxidation-specific epitopes). Distribution of small LDL-IgG-IC in LDL-subfractions isolated from 11 CAD patients. ... Fluorescence counts (1 μg OB/04 antibody per well) were recorded after incubation with europium labelled goat anti-mouse IgG (#M-8770, Sigma Immunochemicals, St. Louis; USA).

    Article Title: A small heat shock/?-crystallin protein from encysted Artemia embryos suppresses tubulin denaturation
    Article Snippet: Paragraph title: Electrophoresis, Western blotting, and protein immunodetection ... Membranes were washed and then incubated for 15 minutes at room temperature, with peroxidase-conjugated secondary antibodies diluted in 10 mM Tris-HCl, 1 M NaCl, 0.5% (v/v) Tween 20, pH 7.4 (HST), including goat anti-rabbit IgG (Bio/Can Scientific, Mississauga, Ontario, Canada), goat anti-mouse IgG (Sigma) and mouse anti-rabbit IgG, and γ-chain–specific clone RG-96 (Sigma), which recognizes only the heavy chain of IgG.

    Laser-Scanning Microscopy:

    Article Title: Exendin-4 Ameliorates Lipotoxicity-induced Glomerular Endothelial Cell Injury by Improving ABC Transporter A1-mediated Cholesterol Efflux in Diabetic apoE Knockout Mice *
    Article Snippet: The sections were then stained with FITC-labeled rabbit anti-goat immunoglobulin (Dako), Tetramethylrhodamine isothiocyanate-labeled rabbit anti-mouse immunoglobulin (Dako), FITC-stained swine anti-rabbit immunoglobulin (Dako), or goat anti-mouse IgG (heavy and light chain) (Millipore). .. The tissue sections were analyzed with a Nikon A1Si confocal laser scanning microscope and NIS-Elements Advanced Research software, version 4.0 (Nikon, Tokyo), and S Plan Fluor ELWD 40× DIC.

    Immunoprecipitation:

    Article Title: Extensive Post-translational Modification of Active and Inactivated Forms of Endogenous p53 *
    Article Snippet: .. Immunoprecipitation efficiency and p53 protein concentration were assessed via immunoblotting as described previously ( ). p53 was visualized with mouse monoclonal antibody D01-HRP (sc-126, Santa Cruz Biotechnologies, Santa Cruz, CA), actin with mouse monoclonal antibody AC-15 HRP (Abcam, Cambridge, MA), and IgG heavy chain with goat anti-mouse HRP (AP124P, Chemicon, EMD Millipore, Billerica, MA). ..

    Marker:

    Article Title: Exendin-4 Ameliorates Lipotoxicity-induced Glomerular Endothelial Cell Injury by Improving ABC Transporter A1-mediated Cholesterol Efflux in Diabetic apoE Knockout Mice *
    Article Snippet: Frozen kidney tissue sections (3 μm) were stained with the primary antibodies mouse anti-GLP-1R (1:200, Abcam), mouse anti-CaMKK (1:100, Abcam), and rabbit anti-ABCA1 (1:200, Abcam) along with the endothelial cell marker mouse anti-CD31 (1:100, Abcam) or rabbit anti-CD31 (Bioworld Technology, Inc.) for 2 h at 37 °C. .. The sections were then stained with FITC-labeled rabbit anti-goat immunoglobulin (Dako), Tetramethylrhodamine isothiocyanate-labeled rabbit anti-mouse immunoglobulin (Dako), FITC-stained swine anti-rabbit immunoglobulin (Dako), or goat anti-mouse IgG (heavy and light chain) (Millipore).

    Lysis:

    Article Title: SHCBP1 regulates apoptosis in lung cancer cells through phosphatase and tensin homolog
    Article Snippet: Protein samples (70 µg) were extracted with lysis buffer (Beyotime Institute of Biotechnology) supplemented with 1% protease inhibitor solution. .. Following washing, the membrane was incubated with secondary anti-rabbit immunoglobulin G (IgG; 1:1,000; cat. no. A3687; Sigma-Aldrich; Merck KGaA) and anti-mouse IgG (1:1,000; cat. no. M8770; Sigma-Aldrich; Merck KGaA) antibodies at room temperature for 1 h. Images were captured on an Odyssey CLx Infrared Imaging system (LI-COR Biosciences).

    Article Title: Extensive Post-translational Modification of Active and Inactivated Forms of Endogenous p53 *
    Article Snippet: Immunoprecipitation efficiency and p53 protein concentration were assessed via immunoblotting as described previously ( ). p53 was visualized with mouse monoclonal antibody D01-HRP (sc-126, Santa Cruz Biotechnologies, Santa Cruz, CA), actin with mouse monoclonal antibody AC-15 HRP (Abcam, Cambridge, MA), and IgG heavy chain with goat anti-mouse HRP (AP124P, Chemicon, EMD Millipore, Billerica, MA). .. Cells were maintained in the presence of etoposide for 44 h, washed twice in PBS, harvested, and incubated in lysis buffer containing N -lauryl sarcosine for analysis via immunoblotting.

    Hood:

    Article Title: Tracking the Emerging Human Pathogen Pseudallescheria boydii by Using Highly Specific Monoclonal Antibodies ▿
    Article Snippet: Wells were washed three times (5 min each time) with PBST, washed once with PBS and once with dH2 O, and air dried at 23°C with a laminar flow hood. .. Antigen extracts prepared in PBST as described above were incubated in the antibody-coated wells for 2 h. The wells were given four 5-min rinses with PBST and then incubated sequentially with HG12 MAb (IgG1) supernatant for 1 h, followed by goat anti-mouse IgG (γ-chain specific) peroxidase conjugate (Sigma), diluted 1 in 1,000 in PBST, for a further hour.

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  • 88
    Millipore rabbit monoclonal anti inos
    <t>NPY</t> inhibits inducible nitric-oxide synthase expression. Microglial cells were treated with 1 μ m NPY and challenged with 100 ng/ml LPS for 6 h to assess the effect of NPY on <t>iNOS</t> (130 kDa) protein levels. A , NPY significantly inhibited LPS-stimulated
    Rabbit Monoclonal Anti Inos, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti inos/product/Millipore
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit monoclonal anti inos - by Bioz Stars, 2020-02
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    82
    Millipore goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate
    Virion-specific <t>IgG</t> and neutralizing-antibody responses in the DNA primed/FI-MCMV-boosted or live virus-vaccinated mice. Mice from each group vaccinated with either pc3-Ua plus PBS plus alum (two mice per group), All-U pDNA plus FI+alum (six mice
    Goat Anti Mouse Immunoglobulin G Igg Horseradish Peroxidase Conjugate, supplied by Millipore, used in various techniques. Bioz Stars score: 82/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate/product/Millipore
    Average 82 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse immunoglobulin g igg horseradish peroxidase conjugate - by Bioz Stars, 2020-02
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    90
    Millipore mouse anti bax monoclonal
    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of <t>pro-caspase-8/9/3</t> (A) and <t>Bcl-2/Bax</t> (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P
    Mouse Anti Bax Monoclonal, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti bax monoclonal/product/Millipore
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    mouse anti bax monoclonal - by Bioz Stars, 2020-02
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    Image Search Results


    NPY inhibits inducible nitric-oxide synthase expression. Microglial cells were treated with 1 μ m NPY and challenged with 100 ng/ml LPS for 6 h to assess the effect of NPY on iNOS (130 kDa) protein levels. A , NPY significantly inhibited LPS-stimulated

    Journal: The Journal of Biological Chemistry

    Article Title: Neuropeptide Y Modulation of Interleukin-1? (IL-1?)-induced Nitric Oxide Production in Microglia *

    doi: 10.1074/jbc.M110.164020

    Figure Lengend Snippet: NPY inhibits inducible nitric-oxide synthase expression. Microglial cells were treated with 1 μ m NPY and challenged with 100 ng/ml LPS for 6 h to assess the effect of NPY on iNOS (130 kDa) protein levels. A , NPY significantly inhibited LPS-stimulated

    Article Snippet: Antibodies used were as follows: rabbit polyclonal anti-NPY (1:1000) (Sigma); sheep polyclonal anti-Y1 R (1:1000) (AbD Serotec, Oxfordshire, UK); rabbit monoclonal anti-iNOS (1:250) (Millipore Corp., Bedford, MA); rat monoclonal anti-CD11b (1:1000) (AbD Serotec); rabbit monoclonal anti-NF-κB p65 (1:100) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) in 0.1% Triton X-100, 0.3% BSA solution; and Alexa Fluor 594 goat anti-rabbit, Alexa Fluor 594 donkey anti-sheep, Alexa Fluor 594 goat anti-rat, Alexa Fluor 488 donkey anti-rabbit, and Alexa Fluor 488 goat anti-rat (all 1:200 in PBS, from Molecular Probes, Eugene, OR).

    Techniques: Expressing

    NPY inhibits IL-1β-induced iNOS protein levels. A , confocal microscopy photomicrographs illustrate microglial cells treated with 1 μ m NPY and 1.5 ng/ml IL-1β for 6 h to assess the role of NPY and IL-1β in iNOS synthesis.

    Journal: The Journal of Biological Chemistry

    Article Title: Neuropeptide Y Modulation of Interleukin-1? (IL-1?)-induced Nitric Oxide Production in Microglia *

    doi: 10.1074/jbc.M110.164020

    Figure Lengend Snippet: NPY inhibits IL-1β-induced iNOS protein levels. A , confocal microscopy photomicrographs illustrate microglial cells treated with 1 μ m NPY and 1.5 ng/ml IL-1β for 6 h to assess the role of NPY and IL-1β in iNOS synthesis.

    Article Snippet: Antibodies used were as follows: rabbit polyclonal anti-NPY (1:1000) (Sigma); sheep polyclonal anti-Y1 R (1:1000) (AbD Serotec, Oxfordshire, UK); rabbit monoclonal anti-iNOS (1:250) (Millipore Corp., Bedford, MA); rat monoclonal anti-CD11b (1:1000) (AbD Serotec); rabbit monoclonal anti-NF-κB p65 (1:100) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) in 0.1% Triton X-100, 0.3% BSA solution; and Alexa Fluor 594 goat anti-rabbit, Alexa Fluor 594 donkey anti-sheep, Alexa Fluor 594 goat anti-rat, Alexa Fluor 488 donkey anti-rabbit, and Alexa Fluor 488 goat anti-rat (all 1:200 in PBS, from Molecular Probes, Eugene, OR).

    Techniques: Confocal Microscopy

    Virion-specific IgG and neutralizing-antibody responses in the DNA primed/FI-MCMV-boosted or live virus-vaccinated mice. Mice from each group vaccinated with either pc3-Ua plus PBS plus alum (two mice per group), All-U pDNA plus FI+alum (six mice

    Journal:

    Article Title: Systemic Priming-Boosting Immunization with a Trivalent Plasmid DNA and Inactivated Murine Cytomegalovirus (MCMV) Vaccine Provides Long-Term Protection against Viral Replication following Systemic or Mucosal MCMV Challenge

    doi: 10.1128/JVI.79.1.159-175.2005

    Figure Lengend Snippet: Virion-specific IgG and neutralizing-antibody responses in the DNA primed/FI-MCMV-boosted or live virus-vaccinated mice. Mice from each group vaccinated with either pc3-Ua plus PBS plus alum (two mice per group), All-U pDNA plus FI+alum (six mice

    Article Snippet: To detect gB, blocked blots were incubated with the monoclonal antibody 2E8.12A (a gift from Lambert Loh, University of Saskatchewan, Saskatoon, Canada), and bound antibody was detected using a goat-anti-mouse immunoglobulin G (IgG) horseradish peroxidase conjugate (Calbiochem) and enhanced chemiluminescence (Supersignal West Pico; Pierce).

    Techniques: Mouse Assay

    Virion-specific IgG and neutralizing-antibody responses in vaccinated mice. On the weeks of the experiment shown in Fig. , four to eight mice per vaccine group were retroorbitally bled and sera were prepared. Arrows and numbers indicate

    Journal:

    Article Title: Systemic Priming-Boosting Immunization with a Trivalent Plasmid DNA and Inactivated Murine Cytomegalovirus (MCMV) Vaccine Provides Long-Term Protection against Viral Replication following Systemic or Mucosal MCMV Challenge

    doi: 10.1128/JVI.79.1.159-175.2005

    Figure Lengend Snippet: Virion-specific IgG and neutralizing-antibody responses in vaccinated mice. On the weeks of the experiment shown in Fig. , four to eight mice per vaccine group were retroorbitally bled and sera were prepared. Arrows and numbers indicate

    Article Snippet: To detect gB, blocked blots were incubated with the monoclonal antibody 2E8.12A (a gift from Lambert Loh, University of Saskatchewan, Saskatoon, Canada), and bound antibody was detected using a goat-anti-mouse immunoglobulin G (IgG) horseradish peroxidase conjugate (Calbiochem) and enhanced chemiluminescence (Supersignal West Pico; Pierce).

    Techniques: Mouse Assay

    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Cell Culture, Transfection, Western Blot, Positive Control

    Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Activation Assay, Expressing, Cell Culture, Transfection, Western Blot

    Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Western Blot