peroxidase conjugated anti rabbit  (Thermo Fisher)


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  • 99
    Name:
    Rabbit anti Rat IgG IgM IgA Peroxidase
    Description:

    Catalog Number:
    sa1-36149
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    None
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    Structured Review

    Thermo Fisher peroxidase conjugated anti rabbit

    https://www.bioz.com/result/peroxidase conjugated anti rabbit/product/Thermo Fisher
    Average 99 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    peroxidase conjugated anti rabbit - by Bioz Stars, 2021-01
    99/100 stars

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    Purification:

    Article Title: Signaling through regulated transcription factor interaction: mapping of a regulatory interaction domain in the Myb-related Bas1p
    Article Snippet: .. Western blot analysis was performed with the ECL Plus™ western blot kit (Amersham) with purified anti-Bas1p (1 µg purified IgG/ml) as primary antibody and peroxidase-conjugated anti-rabbit IgG (diluted 1:6000; Pierce) as secondary antibody. .. From studies of reporter activation using lexA–Bas1p and lexA–Bas2p fusions, Zhang et al . proposed a model for adenine repression of ADE genes in S.cerevisiae stating that complex formation between the two transcription factors Bas1p and Bas2p is a critical regulated step in this response ( ).

    Incubation:

    Article Title: Identification of di-substituted ureas that prevent growth of trypanosomes through inhibition of translation initiation
    Article Snippet: .. The primary antibody was washed 3 times in TBS-T for 10 min and incubated for 1 hour with anti-rabbit IgG peroxidase-conjugated antibody (Thermo Fisher Scientific) diluted 1:20000 in TBS. .. Membranes were washed three times in TBS-T for 10 min and bound antibodies detected by ECL (EMD Millipore) by using an Odyssey Fc System (LI-COR Biosciences).

    Article Title: Knockdown of GSK3β increases basal autophagy and AMPK signalling in nutrient-laden human aortic endothelial cells
    Article Snippet: .. Following washing in TBST, membranes were incubated in peroxidase conjugated anti-rabbit antibody and bands were visualized on film using SuperSignalWest Pico (catalogue # 1856135 and 1856136) or Femto (catalogue # 34095) chemiluminescent substrate (Thermo Scientific). .. Densitometric analyses were carried out using Scion Image software and presented after adjusting for loading controls, GAPDH and β-actin, as used previously [ ].

    Article Title: Profiling and bioinformatic analysis of circular RNA expression regulated by c-Myc
    Article Snippet: .. After washed with TBST, membrane was incubated in horseradish peroxidase-conjugated anti-rabbit antibody for 1 h, and developed by SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific). .. RNA isolation, RNA labeling and RNA hybridization Cell lines were subjected to standard TRIzol RNA isolation (Life Technologies, Carlsbad, CA, USA).

    Article Title: Mucosal immunoglobulins protect the olfactory organ of teleost fish against parasitic infection
    Article Snippet: .. Thereafter, the membranes were blocked with 8% skim milk and incubated with anti-trout IgT (rabbit pAb), anti-trout IgM (mouse monoclonal antibody (mAb)) or biotinylated anti-trout IgD (mouse mAb) antibodies followed by incubation with peroxidase-conjugated anti-rabbit, anti-mouse IgG (Invitrogen) or streptavidin (Invitrogen). .. Immunoreactivity was detected with an enhanced chemiluminescent reagent (Advansta) and scanned by GE Amersham Imager 600 Imaging System (GE Healthcare).

    Lysis:

    Article Title: The LIM-Only Protein PINCH Directly Interacts with Integrin-Linked Kinase and Is Recruited to Integrin-Rich Sites in Spreading Cells
    Article Snippet: .. After five washes with lysis buffer, human ILK associated with MBP-PINCH was detected by immunoblotting with an affinity-purified rabbit polyclonal anti-human ILK antibody (91-5; 69 ng/ml), a horseradish peroxidase-conjugated anti-rabbit immunoglobulin G (IgG) antibody (27 ng/ml), and the SuperSignal chemiluminescent substrate (Pierce). .. A solid-phase-based binding assay was used to detect direct interactions between PINCH and ILK.

    Western Blot:

    Article Title: Signaling through regulated transcription factor interaction: mapping of a regulatory interaction domain in the Myb-related Bas1p
    Article Snippet: .. Western blot analysis was performed with the ECL Plus™ western blot kit (Amersham) with purified anti-Bas1p (1 µg purified IgG/ml) as primary antibody and peroxidase-conjugated anti-rabbit IgG (diluted 1:6000; Pierce) as secondary antibody. .. From studies of reporter activation using lexA–Bas1p and lexA–Bas2p fusions, Zhang et al . proposed a model for adenine repression of ADE genes in S.cerevisiae stating that complex formation between the two transcription factors Bas1p and Bas2p is a critical regulated step in this response ( ).

    Affinity Purification:

    Article Title: The LIM-Only Protein PINCH Directly Interacts with Integrin-Linked Kinase and Is Recruited to Integrin-Rich Sites in Spreading Cells
    Article Snippet: .. After five washes with lysis buffer, human ILK associated with MBP-PINCH was detected by immunoblotting with an affinity-purified rabbit polyclonal anti-human ILK antibody (91-5; 69 ng/ml), a horseradish peroxidase-conjugated anti-rabbit immunoglobulin G (IgG) antibody (27 ng/ml), and the SuperSignal chemiluminescent substrate (Pierce). .. A solid-phase-based binding assay was used to detect direct interactions between PINCH and ILK.

    SDS Page:

    Article Title: Manipulation of Cell Cycle and Chromatin Configuration by Means of Cell-Penetrating Geminin
    Article Snippet: .. Proteins were separated by means of SDS-PAGE, transferred to Immobilon-P (Merck-Millipore, Billerica, MA), immunoblotted with primary antibodies, and visualized with a horseradish peroxidase-conjugated anti-rabbit IgG antibody and SuperSignal West Femto Maximum Sensitivity substrate (Thermo Fisher Scientific) [ ]. .. Cell cycle analysis NIH 3T3 and MEF cells were pulse-labeled with 10 μM 5-Bromo-2'-deoxyuridine (BrdU) for 45 minutes.

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  • 88
    Thermo Fisher rabbit anti goat igg hrp conjugated antibody
    The immunized mouse serum <t>IgG</t> (H+L) titer and IgG subtypes were detected using the indirect ELISA assay. (A) IgG titers of BALB/c mice immunized with rEF-Tu, rHSP70, Mo extracts and PBS sera of each groups were collected on days 3, 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 DAI. (B) Determination of IgG subtypes in sera of the immunized mice. The sera of each group were collected at 35 DAI. ELISA plates were coated with purified rEF-Tu proteins or rHSP70 proteins or Mo extracts of M . ovipneumoniae wild strain Mo-1 at a concentration of 100 ng per well. Anti-mouse IgG (H+L) or IgG1 or IgG2a <t>HRP-conjugated</t> antibodies were used as secondary antibodies. Asterisks indicates the results of the One-Way ANOVA using the Tukey test, compared with the PBS negative control group, with P
    Rabbit Anti Goat Igg Hrp Conjugated Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti goat igg hrp conjugated antibody/product/Thermo Fisher
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
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    90
    Thermo Fisher goat anti rabbit peroxidase conjugated igg abs
    Inhibition of anti-OSs <t>IgG1</t> antibodies (Abs) in mice and rabbit immune sera with OS ligands and CP. (A – C) Inhibition of IgG1 Abs in the pooled sera of BALB/c mice (n = 6 for each glycoconjugate) (A) Di–BSA conjugate antiserum in 1:500 dilution was tested against di–biotin capture material. (B) Tri–BSA conjugate antiserum in 1:500 dilution was tested against tri–biotin capture material. (C) Tetra–BSA conjugate antiserum in 1:1,000 dilution was tested against tetra–biotin capture material. (D – F) Inhibition of <t>IgG</t> Abs in the pooled sera of rabbits (n = 2) immunized with inactive S. pneumoniae type 3 cells. Dilution of sera tested against di–biotin (D) and tri–biotin coating material (E) was 1:500; tetra–biotin coating material (F) was 1:1,000, three assays per antiserum. Different antisera dilutions were used to reach approximately equal antibody level in tested sera samples. IC 50 —the half maximal inhibitory concentration.
    Goat Anti Rabbit Peroxidase Conjugated Igg Abs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 192 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit peroxidase conjugated igg abs/product/Thermo Fisher
    Average 90 stars, based on 192 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit peroxidase conjugated igg abs - by Bioz Stars, 2021-01
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    91
    Thermo Fisher rabbit anti mouse horseradish peroxidase hrp conjugated secondary antibody
    c-di-GMP regulates TfoY protein levels. (A) Western blot of PAGE-separated protein isolated from V. cholerae parent strains carrying either the P 1234 -TL-FULL <t>gfp</t> reporter fusion or the G20U variant. Blots were hybridized with anti-GFP primary antibody and <t>HRP-conjugated</t> secondary antibody and detected using ECL. The blot shown is representative of multiple experiments. (B) Quantification of TfoY-GFP fusion protein present in the lanes depicted in panel A ( n = 3 or 4). Relative amounts of protein were quantified for each lane and normalized to the WT Vc2 riboswitch with the P tac -qrgB * group for each blot (set equal to 1 relative unit of protein). L, low; I, intermediate; H, high. Error bars for all graphs indicate the standard deviation. *, P
    Rabbit Anti Mouse Horseradish Peroxidase Hrp Conjugated Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti mouse horseradish peroxidase hrp conjugated secondary antibody/product/Thermo Fisher
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti mouse horseradish peroxidase hrp conjugated secondary antibody - by Bioz Stars, 2021-01
    91/100 stars
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    The immunized mouse serum IgG (H+L) titer and IgG subtypes were detected using the indirect ELISA assay. (A) IgG titers of BALB/c mice immunized with rEF-Tu, rHSP70, Mo extracts and PBS sera of each groups were collected on days 3, 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 DAI. (B) Determination of IgG subtypes in sera of the immunized mice. The sera of each group were collected at 35 DAI. ELISA plates were coated with purified rEF-Tu proteins or rHSP70 proteins or Mo extracts of M . ovipneumoniae wild strain Mo-1 at a concentration of 100 ng per well. Anti-mouse IgG (H+L) or IgG1 or IgG2a HRP-conjugated antibodies were used as secondary antibodies. Asterisks indicates the results of the One-Way ANOVA using the Tukey test, compared with the PBS negative control group, with P

    Journal: PLoS ONE

    Article Title: Elongation Factor Tu and Heat Shock Protein 70 Are Membrane-Associated Proteins from Mycoplasma ovipneumoniae Capable of Inducing Strong Immune Response in Mice

    doi: 10.1371/journal.pone.0161170

    Figure Lengend Snippet: The immunized mouse serum IgG (H+L) titer and IgG subtypes were detected using the indirect ELISA assay. (A) IgG titers of BALB/c mice immunized with rEF-Tu, rHSP70, Mo extracts and PBS sera of each groups were collected on days 3, 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 DAI. (B) Determination of IgG subtypes in sera of the immunized mice. The sera of each group were collected at 35 DAI. ELISA plates were coated with purified rEF-Tu proteins or rHSP70 proteins or Mo extracts of M . ovipneumoniae wild strain Mo-1 at a concentration of 100 ng per well. Anti-mouse IgG (H+L) or IgG1 or IgG2a HRP-conjugated antibodies were used as secondary antibodies. Asterisks indicates the results of the One-Way ANOVA using the Tukey test, compared with the PBS negative control group, with P

    Article Snippet: Following washing with PBST three times, membranes were incubated with rabbit anti-goat IgG HRP- conjugated antibody at 37°C for 1 h and exposed to SuperSignal® West Pico Chemiluminescent Substrate reagent for detection (Thermo Scientific, USA).

    Techniques: Indirect ELISA, Mouse Assay, Enzyme-linked Immunosorbent Assay, Purification, Concentration Assay, Negative Control

    Inhibition of anti-OSs IgG1 antibodies (Abs) in mice and rabbit immune sera with OS ligands and CP. (A – C) Inhibition of IgG1 Abs in the pooled sera of BALB/c mice (n = 6 for each glycoconjugate) (A) Di–BSA conjugate antiserum in 1:500 dilution was tested against di–biotin capture material. (B) Tri–BSA conjugate antiserum in 1:500 dilution was tested against tri–biotin capture material. (C) Tetra–BSA conjugate antiserum in 1:1,000 dilution was tested against tetra–biotin capture material. (D – F) Inhibition of IgG Abs in the pooled sera of rabbits (n = 2) immunized with inactive S. pneumoniae type 3 cells. Dilution of sera tested against di–biotin (D) and tri–biotin coating material (E) was 1:500; tetra–biotin coating material (F) was 1:1,000, three assays per antiserum. Different antisera dilutions were used to reach approximately equal antibody level in tested sera samples. IC 50 —the half maximal inhibitory concentration.

    Journal: Frontiers in Immunology

    Article Title: Higher Cytokine and Opsonizing Antibody Production Induced by Bovine Serum Albumin (BSA)-Conjugated Tetrasaccharide Related to Streptococcus pneumoniae Type 3 Capsular Polysaccharide

    doi: 10.3389/fimmu.2020.578019

    Figure Lengend Snippet: Inhibition of anti-OSs IgG1 antibodies (Abs) in mice and rabbit immune sera with OS ligands and CP. (A – C) Inhibition of IgG1 Abs in the pooled sera of BALB/c mice (n = 6 for each glycoconjugate) (A) Di–BSA conjugate antiserum in 1:500 dilution was tested against di–biotin capture material. (B) Tri–BSA conjugate antiserum in 1:500 dilution was tested against tri–biotin capture material. (C) Tetra–BSA conjugate antiserum in 1:1,000 dilution was tested against tetra–biotin capture material. (D – F) Inhibition of IgG Abs in the pooled sera of rabbits (n = 2) immunized with inactive S. pneumoniae type 3 cells. Dilution of sera tested against di–biotin (D) and tri–biotin coating material (E) was 1:500; tetra–biotin coating material (F) was 1:1,000, three assays per antiserum. Different antisera dilutions were used to reach approximately equal antibody level in tested sera samples. IC 50 —the half maximal inhibitory concentration.

    Article Snippet: Next, working dilutions of peroxidase-conjugated rabbit anti-mouse IgG1 Abs (Rockland Immunochemicals) or goat anti-rabbit peroxidase-conjugated IgG Abs (Thermo Fisher Scientific) were added, as appropriate.

    Techniques: Inhibition, Mouse Assay, Concentration Assay

    Inhibition of IgG1 antibodies (Abs) recognizing CP of S. pneumoniae type 3 as a coating antigen with OS ligands and CP in immune sera. The horizontal line indicates IC 50 at the point of intersection of the inhibition curves. (A) Inhibition of IgG1 Abs in di–BSA conjugate sera in 1:400 dilution. (B) Inhibition of IgG1 Abs in tri–BSA conjugate sera in 1:800 dilution. (C) Inhibition of IgG1 Abs in tetra–BSA conjugate sera in 1:3,000 dilution. (D) Inhibition of IgG1 Abs in the pooled sera of BALB/c mice (n = 6) immunized with Prevnar 13 and Streptococcus pneumoniae type 3 CP at 1:400 dilution. (E) Inhibition of IgG Abs in the pooled sera of rabbits (n = 2) immunized with inactive S. pneumoniae type 3 cells at 1:2,500 dilution, three assays per antiserum. Different antisera dilutions were used to reach approximately equal antibody level in tested sera samples. IC 50 — the half maximal inhibitory concentration.

    Journal: Frontiers in Immunology

    Article Title: Higher Cytokine and Opsonizing Antibody Production Induced by Bovine Serum Albumin (BSA)-Conjugated Tetrasaccharide Related to Streptococcus pneumoniae Type 3 Capsular Polysaccharide

    doi: 10.3389/fimmu.2020.578019

    Figure Lengend Snippet: Inhibition of IgG1 antibodies (Abs) recognizing CP of S. pneumoniae type 3 as a coating antigen with OS ligands and CP in immune sera. The horizontal line indicates IC 50 at the point of intersection of the inhibition curves. (A) Inhibition of IgG1 Abs in di–BSA conjugate sera in 1:400 dilution. (B) Inhibition of IgG1 Abs in tri–BSA conjugate sera in 1:800 dilution. (C) Inhibition of IgG1 Abs in tetra–BSA conjugate sera in 1:3,000 dilution. (D) Inhibition of IgG1 Abs in the pooled sera of BALB/c mice (n = 6) immunized with Prevnar 13 and Streptococcus pneumoniae type 3 CP at 1:400 dilution. (E) Inhibition of IgG Abs in the pooled sera of rabbits (n = 2) immunized with inactive S. pneumoniae type 3 cells at 1:2,500 dilution, three assays per antiserum. Different antisera dilutions were used to reach approximately equal antibody level in tested sera samples. IC 50 — the half maximal inhibitory concentration.

    Article Snippet: Next, working dilutions of peroxidase-conjugated rabbit anti-mouse IgG1 Abs (Rockland Immunochemicals) or goat anti-rabbit peroxidase-conjugated IgG Abs (Thermo Fisher Scientific) were added, as appropriate.

    Techniques: Inhibition, Mouse Assay, Concentration Assay

    Anti-OS antibody (Ab) classes in pooled sera of BALB/C mice (n = 6) immunized with OS–BSA and CP–CRM 197 conjugates. The biotinylated OSs (di–, tri–, and tetra–biotin) were applied as coating antigens. (A) IgM anti-OS Abs. (B) IgG1 anti-OS Abs. (C) IgG2a anti-OS Abs. (D) IgG2b anti-OS Abs. IgG3 level was low (

    Journal: Frontiers in Immunology

    Article Title: Higher Cytokine and Opsonizing Antibody Production Induced by Bovine Serum Albumin (BSA)-Conjugated Tetrasaccharide Related to Streptococcus pneumoniae Type 3 Capsular Polysaccharide

    doi: 10.3389/fimmu.2020.578019

    Figure Lengend Snippet: Anti-OS antibody (Ab) classes in pooled sera of BALB/C mice (n = 6) immunized with OS–BSA and CP–CRM 197 conjugates. The biotinylated OSs (di–, tri–, and tetra–biotin) were applied as coating antigens. (A) IgM anti-OS Abs. (B) IgG1 anti-OS Abs. (C) IgG2a anti-OS Abs. (D) IgG2b anti-OS Abs. IgG3 level was low (

    Article Snippet: Next, working dilutions of peroxidase-conjugated rabbit anti-mouse IgG1 Abs (Rockland Immunochemicals) or goat anti-rabbit peroxidase-conjugated IgG Abs (Thermo Fisher Scientific) were added, as appropriate.

    Techniques: Mouse Assay

    Anti-CP IgG1 antibody (Ab) titers in BALB/c mice (n = 6) immunized intraperitoneally with di–BSA (A) , tri–BSA (B) , and tetra–BSA (C) conjugates adjuvanted with aluminum hydroxide. Controls: naïve mice (n = 18); mice (n = 6) injected with aluminum hydroxide (250 µg) diluted in saline (three assays per each sera); mice (n = 6) received CP–CRM 197 conjugate, 1.1 µg (three assays per each sera). Data represent individual anti-CP IgG1 Ab titers induced by glycoconjugates, bars indicate median ± SD. Mann–Whitney Rank Sum tests used to evaluate significance between the titer of Ab in anti-tetra–BSA compared with di– and tri–BSA antisera obtained from the mice immunized with 20 µg of the neoglycoconjugates, * P

    Journal: Frontiers in Immunology

    Article Title: Higher Cytokine and Opsonizing Antibody Production Induced by Bovine Serum Albumin (BSA)-Conjugated Tetrasaccharide Related to Streptococcus pneumoniae Type 3 Capsular Polysaccharide

    doi: 10.3389/fimmu.2020.578019

    Figure Lengend Snippet: Anti-CP IgG1 antibody (Ab) titers in BALB/c mice (n = 6) immunized intraperitoneally with di–BSA (A) , tri–BSA (B) , and tetra–BSA (C) conjugates adjuvanted with aluminum hydroxide. Controls: naïve mice (n = 18); mice (n = 6) injected with aluminum hydroxide (250 µg) diluted in saline (three assays per each sera); mice (n = 6) received CP–CRM 197 conjugate, 1.1 µg (three assays per each sera). Data represent individual anti-CP IgG1 Ab titers induced by glycoconjugates, bars indicate median ± SD. Mann–Whitney Rank Sum tests used to evaluate significance between the titer of Ab in anti-tetra–BSA compared with di– and tri–BSA antisera obtained from the mice immunized with 20 µg of the neoglycoconjugates, * P

    Article Snippet: Next, working dilutions of peroxidase-conjugated rabbit anti-mouse IgG1 Abs (Rockland Immunochemicals) or goat anti-rabbit peroxidase-conjugated IgG Abs (Thermo Fisher Scientific) were added, as appropriate.

    Techniques: Mouse Assay, Injection, MANN-WHITNEY

    SB203580, an RIP2 inhibitor downstream of nucleotide-binding oligomerization domain (NOD)2, attenuates CLP-induced sepsis. (A) Peritoneal cells of WT mice were cultured with SB203580 and/or MDP for 24 h, and IL-1β and IL-10 concentrations were measured in culture fractions. (B) Molecules related to NOD2-mediated signal transduction were blotted using peritoneal cells obtained from WT and Nod2 −/− mice injected with SB203580 or PBS 24 h after CLP. (C) Serum and peritoneal IL-1β, IL-10, and C5a levels were estimated in WT (n = 4) and Nod2 −/− (n = 3) mice injected with SB203580 (n = 4 in WT, n = 3 in Nod2 −/− ) or PBS 24 h after CLP by ELISA. (D) The levels of CD55 expression on F4/80 − Ly6-G + cells from WT (n = 3) and WT mice injected with SB203580 (n = 3) were measured 24 h after CLP. (mean fluorescence intensity [MFI] of CD55 expression in the panels, diagrams filled with gray for istotype-matched control IgG, solid lines for CD55) (E) The percentages of surviving mice were estimated during CLP-induced sepsis ( a P = 0.0312, log-rank test, n = 6–8 per group; WT mice injected with SB203580 vs. PBS). *P

    Journal: PLoS Pathogens

    Article Title: NOD2-mediated Suppression of CD55 on Neutrophils Enhances C5a Generation During Polymicrobial Sepsis

    doi: 10.1371/journal.ppat.1003351

    Figure Lengend Snippet: SB203580, an RIP2 inhibitor downstream of nucleotide-binding oligomerization domain (NOD)2, attenuates CLP-induced sepsis. (A) Peritoneal cells of WT mice were cultured with SB203580 and/or MDP for 24 h, and IL-1β and IL-10 concentrations were measured in culture fractions. (B) Molecules related to NOD2-mediated signal transduction were blotted using peritoneal cells obtained from WT and Nod2 −/− mice injected with SB203580 or PBS 24 h after CLP. (C) Serum and peritoneal IL-1β, IL-10, and C5a levels were estimated in WT (n = 4) and Nod2 −/− (n = 3) mice injected with SB203580 (n = 4 in WT, n = 3 in Nod2 −/− ) or PBS 24 h after CLP by ELISA. (D) The levels of CD55 expression on F4/80 − Ly6-G + cells from WT (n = 3) and WT mice injected with SB203580 (n = 3) were measured 24 h after CLP. (mean fluorescence intensity [MFI] of CD55 expression in the panels, diagrams filled with gray for istotype-matched control IgG, solid lines for CD55) (E) The percentages of surviving mice were estimated during CLP-induced sepsis ( a P = 0.0312, log-rank test, n = 6–8 per group; WT mice injected with SB203580 vs. PBS). *P

    Article Snippet: Antibodies against phosphor–p38, p38 (Cell Signaling Technology, MA, USA), phosphor-Rip2 (Thermo scientific, Rockford, USA), Rip2 (Santa Cruz Biotechnology, CA, USA), Bb (Santa Cruz Biotechnology), NOD2 (Santa Cruz Biotechnology), and a horseradish peroxidase-conjugated goat anti-rabbit IgG (Thermo scientific) were used.

    Techniques: Binding Assay, Mouse Assay, Cell Culture, Transduction, Injection, Enzyme-linked Immunosorbent Assay, Expressing, Fluorescence

    IL-1β-dependent IL-10 production mediated by nucleotide-binding oligomerization domain (NOD) 2 enhances C5a generation by suppressing CD55 expression on Ly6-G + cells during sepsis. (A) CD55 and CR1/2 expression on gated F4/80 − Ly6-G + peritoneal cells from WT, Nod2 −/− , and Nod2 −/− mice injected with recombinant IL-1β or IL-10 was estimated 24 h after CLP (mean fluorescence intensity [MFI] of CD55 expression in the panels). (B) CD55 expression on gated F4/80 − Ly6-G + peritoneal cells from Il-10 −/− or Il-10 −/− mice injected with recombinant IL-1β was estimated 24 h after CLP (MFI of CD55 expression in the panels) (C) To block IL-10 receptor engagement in vivo , anti-IL10 receptor mAbs were i.p. injected into WT and Nod2 −/− mice administered recombinant IL-10 during CLP-induced sepsis. CD55 expression on gated F4/80 − Ly6-G + peritoneal cells from these mice 24 h after CLP was evaluated. (D) The levels of CD55 expression on F4/80 − Ly6-G + peritoneal cells were compared in WT, Nod2 −/− , IL-10 −/− , and Il-1r −/− mice 24 h after CLP. (A–D) anti-CD55 mAb (lines) and control IgG (diagrams filled with gray) were used. (E) Peritoneal cells from WT and Nod2 −/− mice 24 h after CLP were blotted for Bb factor. (F) Peritoneal cells from WT and Nod2 −/− mice 12 h after CLP were incubated with RPMI media containing 10% WT mouse serum for 24 h. (G and H) To evaluate the effect of CD55 on C5a generation in vivo , WT, Nod2 −/− , (G) or Nod2 −/− mice given recombinant IL-10 (H) were i.p. injected with soluble CD55 12 h after CLP. Serum and peritoneal C5a levels and the survival percentages of these mice were measured during CLP-induced sepsis ( a P = 0.0124, log-rank test; WT [n = 8], Nod2 −/− [n = 8 ], and soluble CD55-injected WT [n = 6 ] or Nod2 −/− mice [n = 8 ] in G, a P = 0.0024, log-rank test; Nod2 −/− mice [n = 8], Nod2 −/− mice injected with recombinant IL-10 [n = 8] or recombinant IL-10 and soluble CD55 [n = 6] in H). *P

    Journal: PLoS Pathogens

    Article Title: NOD2-mediated Suppression of CD55 on Neutrophils Enhances C5a Generation During Polymicrobial Sepsis

    doi: 10.1371/journal.ppat.1003351

    Figure Lengend Snippet: IL-1β-dependent IL-10 production mediated by nucleotide-binding oligomerization domain (NOD) 2 enhances C5a generation by suppressing CD55 expression on Ly6-G + cells during sepsis. (A) CD55 and CR1/2 expression on gated F4/80 − Ly6-G + peritoneal cells from WT, Nod2 −/− , and Nod2 −/− mice injected with recombinant IL-1β or IL-10 was estimated 24 h after CLP (mean fluorescence intensity [MFI] of CD55 expression in the panels). (B) CD55 expression on gated F4/80 − Ly6-G + peritoneal cells from Il-10 −/− or Il-10 −/− mice injected with recombinant IL-1β was estimated 24 h after CLP (MFI of CD55 expression in the panels) (C) To block IL-10 receptor engagement in vivo , anti-IL10 receptor mAbs were i.p. injected into WT and Nod2 −/− mice administered recombinant IL-10 during CLP-induced sepsis. CD55 expression on gated F4/80 − Ly6-G + peritoneal cells from these mice 24 h after CLP was evaluated. (D) The levels of CD55 expression on F4/80 − Ly6-G + peritoneal cells were compared in WT, Nod2 −/− , IL-10 −/− , and Il-1r −/− mice 24 h after CLP. (A–D) anti-CD55 mAb (lines) and control IgG (diagrams filled with gray) were used. (E) Peritoneal cells from WT and Nod2 −/− mice 24 h after CLP were blotted for Bb factor. (F) Peritoneal cells from WT and Nod2 −/− mice 12 h after CLP were incubated with RPMI media containing 10% WT mouse serum for 24 h. (G and H) To evaluate the effect of CD55 on C5a generation in vivo , WT, Nod2 −/− , (G) or Nod2 −/− mice given recombinant IL-10 (H) were i.p. injected with soluble CD55 12 h after CLP. Serum and peritoneal C5a levels and the survival percentages of these mice were measured during CLP-induced sepsis ( a P = 0.0124, log-rank test; WT [n = 8], Nod2 −/− [n = 8 ], and soluble CD55-injected WT [n = 6 ] or Nod2 −/− mice [n = 8 ] in G, a P = 0.0024, log-rank test; Nod2 −/− mice [n = 8], Nod2 −/− mice injected with recombinant IL-10 [n = 8] or recombinant IL-10 and soluble CD55 [n = 6] in H). *P

    Article Snippet: Antibodies against phosphor–p38, p38 (Cell Signaling Technology, MA, USA), phosphor-Rip2 (Thermo scientific, Rockford, USA), Rip2 (Santa Cruz Biotechnology, CA, USA), Bb (Santa Cruz Biotechnology), NOD2 (Santa Cruz Biotechnology), and a horseradish peroxidase-conjugated goat anti-rabbit IgG (Thermo scientific) were used.

    Techniques: Binding Assay, Expressing, Mouse Assay, Injection, Recombinant, Fluorescence, Blocking Assay, In Vivo, Incubation

    c-di-GMP regulates TfoY protein levels. (A) Western blot of PAGE-separated protein isolated from V. cholerae parent strains carrying either the P 1234 -TL-FULL gfp reporter fusion or the G20U variant. Blots were hybridized with anti-GFP primary antibody and HRP-conjugated secondary antibody and detected using ECL. The blot shown is representative of multiple experiments. (B) Quantification of TfoY-GFP fusion protein present in the lanes depicted in panel A ( n = 3 or 4). Relative amounts of protein were quantified for each lane and normalized to the WT Vc2 riboswitch with the P tac -qrgB * group for each blot (set equal to 1 relative unit of protein). L, low; I, intermediate; H, high. Error bars for all graphs indicate the standard deviation. *, P

    Journal: Journal of Bacteriology

    Article Title: Cyclic di-GMP Regulates TfoY in Vibrio cholerae To Control Motility by both Transcriptional and Posttranscriptional Mechanisms

    doi: 10.1128/JB.00578-17

    Figure Lengend Snippet: c-di-GMP regulates TfoY protein levels. (A) Western blot of PAGE-separated protein isolated from V. cholerae parent strains carrying either the P 1234 -TL-FULL gfp reporter fusion or the G20U variant. Blots were hybridized with anti-GFP primary antibody and HRP-conjugated secondary antibody and detected using ECL. The blot shown is representative of multiple experiments. (B) Quantification of TfoY-GFP fusion protein present in the lanes depicted in panel A ( n = 3 or 4). Relative amounts of protein were quantified for each lane and normalized to the WT Vc2 riboswitch with the P tac -qrgB * group for each blot (set equal to 1 relative unit of protein). L, low; I, intermediate; H, high. Error bars for all graphs indicate the standard deviation. *, P

    Article Snippet: Blots were blocked with 5% nonfat milk in Tris-buffered saline with 0.1% Tween 20 (TBS-T) and hybridized with a mouse anti-GFP primary antibody and a rabbit anti-mouse horseradish peroxidase (HRP)-conjugated secondary antibody (both Thermo Fisher Scientific).

    Techniques: Western Blot, Polyacrylamide Gel Electrophoresis, Isolation, Variant Assay, Standard Deviation