pbs  (Abcam)

 
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    Name:
    Phosphate Buffered Saline PBS Casein ELISA Reagent
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    ab171532
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    Structured Review

    Abcam pbs

    https://www.bioz.com/result/pbs/product/Abcam
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbs - by Bioz Stars, 2021-03
    86/100 stars

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    Related Articles

    Blocking Assay:

    Article Title: Divalent cation-induced conformational changes of influenza virus hemagglutinin
    Article Snippet: The 8-well strip was coated with the native, Zn2+, or acidic-pH incubated CU44 HA in a total volume of 100 µl (2 ng/µl) in microplates (Corning). .. Following blocking with 1% BSA and PBS washing, primary antibodies (the stem-specific CR9114 or head-specific D2 H1-1/H3-1) were applied at 400, 80, 1, 3, 0.64, 0.128, and 0.026 ng per well at 37 °C for 1 h. They were then washed with PBS, and detected by anti-human IgG1-HRP (Abcam, Cambridge, UK) as the secondary antibody for 1 h. 150 µl of 3,3′,5,5′-tetramethylbenzidine (Sigma-Aldrich) was used as substrate and the reaction was stopped by the addition of 2 N sulfuric acid, and the absorbance at 450 nm was monitored. .. DSFDSF was used to measure the shift in transition temperature of HA at different pH conditions, using Stratagene MX3005P (AGILENT TECHNOLOGIES, Santa Clara, CA, USA).

    Article Title: Human Neutrophil Peptide 1 Limits Hypercholesterolemia-induced Atherosclerosis by Increasing Hepatic LDL Clearance
    Article Snippet: .. The membrane was incubated overnight at 4 °C with respective primary antibody (anti-HNP1, anti-ApoA, anti-ApoB, anti ApoC3, 1:5000, all Abcam) in PBS supplemented with Roti® Block Solution. .. The membrane was washed with PBS supplemented with 0.05% Tween® 20 and then incubated with diluted horseradish peroxidase (HRP)-linked secondary antibody (1:5000 dilution; Cell Signaling Technology) in PBS supplemented with Roti® Block Solution for 1 h at room temperature, followed by three additional washes in PBS/Tween for 15 min each (3 × 15 min).

    Article Title: A comparison of BMP2 delivery by coacervate and gene therapy for promoting human muscle-derived stem cell-mediated articular cartilage repair
    Article Snippet: Immunohistochemistry IHC staining of GFP-positive cells was used to reveal donor cells in the regenerated cartilage and Col2 staining was used to detect specific cartilage matrix collagen 2. .. Briefly, after deparaffinization, washing, and blocking with 5% donkey serum in PBS, sections were incubated with rabbit anti-GFP antibody (ab290, Abcam, 1:1000 dilution) and rabbit anti-Col2 (ab34712, Abcam: 1:400 dilution) in 5% donkey serum overnight. .. For Col2 staining, antigen retrieval was performed using 2% hyaluronidase (H3506-5G, Sigma, ) in PBS at room temperature for 30 min, followed by washing with PBS three times before blocking and incubation with primary antibody.

    Incubation:

    Article Title: Human Neutrophil Peptide 1 Limits Hypercholesterolemia-induced Atherosclerosis by Increasing Hepatic LDL Clearance
    Article Snippet: .. The membrane was incubated overnight at 4 °C with respective primary antibody (anti-HNP1, anti-ApoA, anti-ApoB, anti ApoC3, 1:5000, all Abcam) in PBS supplemented with Roti® Block Solution. .. The membrane was washed with PBS supplemented with 0.05% Tween® 20 and then incubated with diluted horseradish peroxidase (HRP)-linked secondary antibody (1:5000 dilution; Cell Signaling Technology) in PBS supplemented with Roti® Block Solution for 1 h at room temperature, followed by three additional washes in PBS/Tween for 15 min each (3 × 15 min).

    Article Title: Cancer-Associated Fibroblasts Promote the Chemo-resistance in Gastric Cancer through Secreting IL-11 Targeting JAK/STAT3/Bcl2 Pathway
    Article Snippet: To detect the expression of JAK and STAT3 in gastric cancer tissues, antigen retrieval was done using citric acid and sodium citrate in a Microwave oven (Media, Guangdong, China). .. Then the sections or gastric cancer cells (fixed with 4% paraformaldehyde and permeabilized with 0.3% Triton X-100) were blocked with 5% BSA in PBS and incubated with p-JAK (1:200, Abcam) or p-STAT3 (1:500, Abcam) at 4°C overnight, followed by secondary antibodies (Thermo Fisher) for 1 hour at room temperature. .. Nuclei were stained with the DAPI solution (1 μg/mL).

    Article Title: Binding of Serum Mannan Binding Lectin to a Cell Integrity-Defective Cryptococcus neoformans ccr4? Mutant ▿
    Article Snippet: Approximately 5 × 106 cells were washed in PBS and incubated for 1 h in 100% human AB serum (Sigma) at 37°C to allow opsonization. .. Cells were washed three times with 1 ml PBS and incubated with either anti-human MBL monoclonal antibody HYB-131-01 (Abcam) or an immunoglobulin G1 (IgG1) isotype control (anti-c- myc clone 9E10; Sigma) at a dilution of 1:250 in PBS with 0.5 mg/ml bovine serum albumin for 1 h at room temperature. .. After three 1-ml washes in PBS, cells were incubated with an AlexaFluor 488-conjugated anti-mouse antibody (Invitrogen) at a 1:500 dilution in PBS-bovine serum albumin for 1 h at room temperature.

    Article Title: A comparison of BMP2 delivery by coacervate and gene therapy for promoting human muscle-derived stem cell-mediated articular cartilage repair
    Article Snippet: Immunohistochemistry IHC staining of GFP-positive cells was used to reveal donor cells in the regenerated cartilage and Col2 staining was used to detect specific cartilage matrix collagen 2. .. Briefly, after deparaffinization, washing, and blocking with 5% donkey serum in PBS, sections were incubated with rabbit anti-GFP antibody (ab290, Abcam, 1:1000 dilution) and rabbit anti-Col2 (ab34712, Abcam: 1:400 dilution) in 5% donkey serum overnight. .. For Col2 staining, antigen retrieval was performed using 2% hyaluronidase (H3506-5G, Sigma, ) in PBS at room temperature for 30 min, followed by washing with PBS three times before blocking and incubation with primary antibody.

    Article Title: Visualization of chikungunya virus infection in vitro and in vivo
    Article Snippet: .. The fixed bone marrow was washed three times with PBS and then incubated with mouse polyclonal anti-CHIKV antibody (1:200 dilution with PBS) for 1 h. After washing with PBS for three times, the bone marrow was incubated with a secondary antibody (goat anti-mouse IgG H & L-Alexa fluor 488 preabsorbed, Abcam, Cat # ab150117; 1:200) at room temperature for 1 h. After three times of washing with PBS, the slide was examined by a fluorescent microscope. .. Plaque assays BHK-21 cells (2 × 105 cells per well in a 12-well plate) were infected with 10-fold serial dilutions of WT or CHIKV-iRFP viruses for 1 h at 37°C and then overlaid with DMEM containing 2% FBS and 2% methyl cellulose.

    Negative Control:

    Article Title: Expression of BDNF, TrkB, VEGF and CD105 is associated with pelvic lymph node metastasis and prognosis in IB2-stage squamous cell carcinoma
    Article Snippet: After washing with PBS, the samples were blocked by incubation with 5% goat serum (cat. no. ab 138478; Abcam) at room temperature for 10 min. .. The primary antibody reaction was performed at 4°C overnight in 1% goat serum in PBS containing the corresponding antibody [BDNF (1:500; cat. no. ab203573; Abcam), TrkB (1:500; cat. no. SC-8316; Santa Cruz Biotechnology, Inc.), VEGF (1:250; cat. no. ab32152; Abcam), CD105 (1:500; cat. no. ab135528; Abcam) and PBS (as the negative control)]. .. After triple washing with PBS, the slides were incubated with the secondary antibody (Goat Anti-Mouse Anti-Rabbit IgG/IgM H & L; 1:100; cat. no. ab2891; Abcam) for 30 min at room temperature.

    Microscopy:

    Article Title: Visualization of chikungunya virus infection in vitro and in vivo
    Article Snippet: .. The fixed bone marrow was washed three times with PBS and then incubated with mouse polyclonal anti-CHIKV antibody (1:200 dilution with PBS) for 1 h. After washing with PBS for three times, the bone marrow was incubated with a secondary antibody (goat anti-mouse IgG H & L-Alexa fluor 488 preabsorbed, Abcam, Cat # ab150117; 1:200) at room temperature for 1 h. After three times of washing with PBS, the slide was examined by a fluorescent microscope. .. Plaque assays BHK-21 cells (2 × 105 cells per well in a 12-well plate) were infected with 10-fold serial dilutions of WT or CHIKV-iRFP viruses for 1 h at 37°C and then overlaid with DMEM containing 2% FBS and 2% methyl cellulose.

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  • 92
    Abcam atp1a2
    Effect of herb pair HZ-GC on protein expression levels of the corresponding candidate targets (A) Adcy2; (B) <t>Atp1a2;</t> (C) Gsr; (D) Iyd; (E) Pdia4; (F) Plcb1; (G) Tg; (H) Tpo according to Western blot analysis Data are represented as the mean ± S.E. ‘*’, ‘**’, and ‘***’, P
    Atp1a2, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pbs  (Abcam)
    99
    Abcam pbs
    Interleukin 11 (IL-11)/IL-11R signaling pathway induced the chemo-resistance through <t>JAK/STAT3</t> pathway. (A) Immunofluoresence of p-JAK2 in BGC823 and SGC7901 cells pre-treated with or without cultured medium of cancer-associated-fibroblasts (CAFs-CM)/rhIL-11 in the presence or absence of IL-11 neutralizing antibody. (B) Immunofluoresence of p-STAT3 in BGC823 and SGC7901 cells pre-treated with or without CAFs-CM/rhIL-11 (10 ng/mL) in the presence or absence of IL-11 neutralizing antibody (25 μg/mL). (C) Western blotting of p-JAK2, total JAK2 and β-actin in BGC823 and SGC7901 cells pre-treated with or without CAFs-CM/rhIL-11 (10 ng/mL) in the presence or absence of IL-11 neutralizing antibody (25 μg/mL). (D) Western blotting of p-STAT3, total STAT3, and β-actin in BGC823 and SGC7901 cells pre-treated with or without CAFs-CM/rhIL-11 (10 ng/mL) in the presence or absence of IL-11 neutralizing antibody (25 μg/mL). (E) The cell viability of BGC823 cells treated with 6 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the presence or absence of ruxolitinib (5 μM). <t>PBS,</t> phosphate buffered saline; Dox, doxorubicin; Eto, etoposide. (F) The cell viability of SGC7901 cells treated with 4 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the presence or absence of ruxolitinib (5 μM). (G) The cell viability of BGC823 cells treated with 6 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the wild type or shSTAT3 cells. (H) The cell viability of SGC7901 cells treated with 4 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the wild type or shSTAT3 cells. The data was presented as the mean±standard error of mean from three independent experiments. ** p
    Pbs, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Abcam anti human ifn β
    Expression of VP4 inhibits TNF-α- or SeV-induced activation of <t>IFN-α,</t> <t>IFN-β,</t> and NF-κB promoters. (A to C) Effects of VP4 on TNF-induced activation of IFN-α, IFN-β, and NF-κB promoters. HEK293T cells
    Anti Human Ifn β, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effect of herb pair HZ-GC on protein expression levels of the corresponding candidate targets (A) Adcy2; (B) Atp1a2; (C) Gsr; (D) Iyd; (E) Pdia4; (F) Plcb1; (G) Tg; (H) Tpo according to Western blot analysis Data are represented as the mean ± S.E. ‘*’, ‘**’, and ‘***’, P

    Journal: Oncotarget

    Article Title: Thyroid hormone synthesis: a potential target of a Chinese herbal formula Haizao Yuhu Decoction acting on iodine-deficient goiter

    doi: 10.18632/oncotarget.10329

    Figure Lengend Snippet: Effect of herb pair HZ-GC on protein expression levels of the corresponding candidate targets (A) Adcy2; (B) Atp1a2; (C) Gsr; (D) Iyd; (E) Pdia4; (F) Plcb1; (G) Tg; (H) Tpo according to Western blot analysis Data are represented as the mean ± S.E. ‘*’, ‘**’, and ‘***’, P

    Article Snippet: Antibodies against the following proteins were used: Adcy1 (rabbit polyclonal antibody, dilution 1:200, Abcam, Cambridge, UK), Adcy2 (rabbit polyclonal antibody, dilution 1:200, Abcam, Cambridge, UK), Atp1a2 (rabbit monoclonal antibody, dilution 1:500, Abcam, Cambridge, UK), Creb1 (rabbit monoclonal antibody, dilution 1:500, Abcam, Cambridge, UK), Gsr (rabbit monoclonal antibody, dilution 1:2000, Abcam, Cambridge, UK), Hspa5 (rabbit monoclonal antibody, dilution 1:2000, Abcam, Cambridge, UK), Lyd (rabbit polyclonal antibody, dilution 1:1000, Abcam, Cambridge, UK), Pdia4 (rabbit polyclonal antibody, dilution 1:1000, Abcam, Cambridge, UK), Plcb1 (rabbit monoclonal antibody, dilution 1:1000, Abcam, Cambridge, UK), Prkca (rabbit monoclonal antibody, dilution 1:2000, Abcam, Cambridge, UK), Prkcb (rabbit monoclonal antibody, dilution 1:2000, Abcam, Cambridge, UK), Tg (rabbit monoclonal antibody, dilution 1:40000, Abcam, Cambridge, UK), and Tpo (goat polyclonal antibody, dilution 1:500, RD, Minnesota, US).

    Techniques: Expressing, Western Blot

    Interleukin 11 (IL-11)/IL-11R signaling pathway induced the chemo-resistance through JAK/STAT3 pathway. (A) Immunofluoresence of p-JAK2 in BGC823 and SGC7901 cells pre-treated with or without cultured medium of cancer-associated-fibroblasts (CAFs-CM)/rhIL-11 in the presence or absence of IL-11 neutralizing antibody. (B) Immunofluoresence of p-STAT3 in BGC823 and SGC7901 cells pre-treated with or without CAFs-CM/rhIL-11 (10 ng/mL) in the presence or absence of IL-11 neutralizing antibody (25 μg/mL). (C) Western blotting of p-JAK2, total JAK2 and β-actin in BGC823 and SGC7901 cells pre-treated with or without CAFs-CM/rhIL-11 (10 ng/mL) in the presence or absence of IL-11 neutralizing antibody (25 μg/mL). (D) Western blotting of p-STAT3, total STAT3, and β-actin in BGC823 and SGC7901 cells pre-treated with or without CAFs-CM/rhIL-11 (10 ng/mL) in the presence or absence of IL-11 neutralizing antibody (25 μg/mL). (E) The cell viability of BGC823 cells treated with 6 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the presence or absence of ruxolitinib (5 μM). PBS, phosphate buffered saline; Dox, doxorubicin; Eto, etoposide. (F) The cell viability of SGC7901 cells treated with 4 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the presence or absence of ruxolitinib (5 μM). (G) The cell viability of BGC823 cells treated with 6 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the wild type or shSTAT3 cells. (H) The cell viability of SGC7901 cells treated with 4 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the wild type or shSTAT3 cells. The data was presented as the mean±standard error of mean from three independent experiments. ** p

    Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

    Article Title: Cancer-Associated Fibroblasts Promote the Chemo-resistance in Gastric Cancer through Secreting IL-11 Targeting JAK/STAT3/Bcl2 Pathway

    doi: 10.4143/crt.2018.031

    Figure Lengend Snippet: Interleukin 11 (IL-11)/IL-11R signaling pathway induced the chemo-resistance through JAK/STAT3 pathway. (A) Immunofluoresence of p-JAK2 in BGC823 and SGC7901 cells pre-treated with or without cultured medium of cancer-associated-fibroblasts (CAFs-CM)/rhIL-11 in the presence or absence of IL-11 neutralizing antibody. (B) Immunofluoresence of p-STAT3 in BGC823 and SGC7901 cells pre-treated with or without CAFs-CM/rhIL-11 (10 ng/mL) in the presence or absence of IL-11 neutralizing antibody (25 μg/mL). (C) Western blotting of p-JAK2, total JAK2 and β-actin in BGC823 and SGC7901 cells pre-treated with or without CAFs-CM/rhIL-11 (10 ng/mL) in the presence or absence of IL-11 neutralizing antibody (25 μg/mL). (D) Western blotting of p-STAT3, total STAT3, and β-actin in BGC823 and SGC7901 cells pre-treated with or without CAFs-CM/rhIL-11 (10 ng/mL) in the presence or absence of IL-11 neutralizing antibody (25 μg/mL). (E) The cell viability of BGC823 cells treated with 6 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the presence or absence of ruxolitinib (5 μM). PBS, phosphate buffered saline; Dox, doxorubicin; Eto, etoposide. (F) The cell viability of SGC7901 cells treated with 4 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the presence or absence of ruxolitinib (5 μM). (G) The cell viability of BGC823 cells treated with 6 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the wild type or shSTAT3 cells. (H) The cell viability of SGC7901 cells treated with 4 μg/mL DDP, 6 μM etoposide, and 6 μM doxorubicin respectively with or without CAFs-CM or rhIL-11 (10 ng/mL) pre-co-cultured in the wild type or shSTAT3 cells. The data was presented as the mean±standard error of mean from three independent experiments. ** p

    Article Snippet: Then the sections or gastric cancer cells (fixed with 4% paraformaldehyde and permeabilized with 0.3% Triton X-100) were blocked with 5% BSA in PBS and incubated with p-JAK (1:200, Abcam) or p-STAT3 (1:500, Abcam) at 4°C overnight, followed by secondary antibodies (Thermo Fisher) for 1 hour at room temperature.

    Techniques: Cell Culture, Western Blot

    Interleukin 11 (IL-11) triggers the JAK/STAT3 pathway to elevate Bcl2 expression. (A) The expression of Bcl2 in BGC823 and SGC7901 cells treated with cultured medium of cancer-associated-fibroblasts (CAFs-CM) or rhIL-11 in the presence or absence of ruxolitinib (5 μM) in mRNA level. (B) The expression of Bcl2 in BGC823 and SGC7901 cells treated with CAFs-CM or rhIL-11 (10 ng/mL) in the presence or absence of ruxolitinib (5 μM) in protein level. (C) The cell viability of BGC823 cells treated with CAFs-CM or rhIL-11 in wild type or Bcl2 silenced cells. (D) The cell viability of SGC7901 cells treated with CAFs-CM or rhIL-11 in wild type or Bcl2 silenced cells. PBS, phosphate buffered saline; Dox, doxorubicin; Eto, etoposide. The data was presented as the mean±standard error of mean from three independent experiments. ** p

    Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

    Article Title: Cancer-Associated Fibroblasts Promote the Chemo-resistance in Gastric Cancer through Secreting IL-11 Targeting JAK/STAT3/Bcl2 Pathway

    doi: 10.4143/crt.2018.031

    Figure Lengend Snippet: Interleukin 11 (IL-11) triggers the JAK/STAT3 pathway to elevate Bcl2 expression. (A) The expression of Bcl2 in BGC823 and SGC7901 cells treated with cultured medium of cancer-associated-fibroblasts (CAFs-CM) or rhIL-11 in the presence or absence of ruxolitinib (5 μM) in mRNA level. (B) The expression of Bcl2 in BGC823 and SGC7901 cells treated with CAFs-CM or rhIL-11 (10 ng/mL) in the presence or absence of ruxolitinib (5 μM) in protein level. (C) The cell viability of BGC823 cells treated with CAFs-CM or rhIL-11 in wild type or Bcl2 silenced cells. (D) The cell viability of SGC7901 cells treated with CAFs-CM or rhIL-11 in wild type or Bcl2 silenced cells. PBS, phosphate buffered saline; Dox, doxorubicin; Eto, etoposide. The data was presented as the mean±standard error of mean from three independent experiments. ** p

    Article Snippet: Then the sections or gastric cancer cells (fixed with 4% paraformaldehyde and permeabilized with 0.3% Triton X-100) were blocked with 5% BSA in PBS and incubated with p-JAK (1:200, Abcam) or p-STAT3 (1:500, Abcam) at 4°C overnight, followed by secondary antibodies (Thermo Fisher) for 1 hour at room temperature.

    Techniques: Expressing, Cell Culture

    Expression of VP4 inhibits TNF-α- or SeV-induced activation of IFN-α, IFN-β, and NF-κB promoters. (A to C) Effects of VP4 on TNF-induced activation of IFN-α, IFN-β, and NF-κB promoters. HEK293T cells

    Journal: Journal of Virology

    Article Title: Critical Roles of Glucocorticoid-Induced Leucine Zipper in Infectious Bursal Disease Virus (IBDV)-Induced Suppression of Type I Interferon Expression and Enhancement of IBDV Growth in Host Cells via Interaction with VP4

    doi: 10.1128/JVI.02421-12

    Figure Lengend Snippet: Expression of VP4 inhibits TNF-α- or SeV-induced activation of IFN-α, IFN-β, and NF-κB promoters. (A to C) Effects of VP4 on TNF-induced activation of IFN-α, IFN-β, and NF-κB promoters. HEK293T cells

    Article Snippet: To neutralize interferon's activity, 2 μg of anti-human IFN-α1 (ab11408; Abcam) and anti-human IFN-β (ab6979; Abcam) were added to GILZ RNAi cell culture or RNAi controls 3 h before IBDV infection.

    Techniques: Expressing, Activation Assay

    Infection of HEK293T cells with IBDV strain Lx inhibits TNF-induced expression of IFN-α, IFN-β, and NF-κB. (A to D) Replication of IBDV strain Lx in HEK293T cells. HEK293T cells were mock infected (A and B) or infected with IBDV

    Journal: Journal of Virology

    Article Title: Critical Roles of Glucocorticoid-Induced Leucine Zipper in Infectious Bursal Disease Virus (IBDV)-Induced Suppression of Type I Interferon Expression and Enhancement of IBDV Growth in Host Cells via Interaction with VP4

    doi: 10.1128/JVI.02421-12

    Figure Lengend Snippet: Infection of HEK293T cells with IBDV strain Lx inhibits TNF-induced expression of IFN-α, IFN-β, and NF-κB. (A to D) Replication of IBDV strain Lx in HEK293T cells. HEK293T cells were mock infected (A and B) or infected with IBDV

    Article Snippet: To neutralize interferon's activity, 2 μg of anti-human IFN-α1 (ab11408; Abcam) and anti-human IFN-β (ab6979; Abcam) were added to GILZ RNAi cell culture or RNAi controls 3 h before IBDV infection.

    Techniques: Infection, Expressing

    GILZ mediates the inhibitory effect of VP4 on poly(I·C)-induced expressions of IFN-α, IFN-β, and NF-κB in DF-1 cells. (A and B) Effect of GILZ RNAi on the expression of endogenous GILZ in DF-1 cells. DF-1 cells (2.0 ×

    Journal: Journal of Virology

    Article Title: Critical Roles of Glucocorticoid-Induced Leucine Zipper in Infectious Bursal Disease Virus (IBDV)-Induced Suppression of Type I Interferon Expression and Enhancement of IBDV Growth in Host Cells via Interaction with VP4

    doi: 10.1128/JVI.02421-12

    Figure Lengend Snippet: GILZ mediates the inhibitory effect of VP4 on poly(I·C)-induced expressions of IFN-α, IFN-β, and NF-κB in DF-1 cells. (A and B) Effect of GILZ RNAi on the expression of endogenous GILZ in DF-1 cells. DF-1 cells (2.0 ×

    Article Snippet: To neutralize interferon's activity, 2 μg of anti-human IFN-α1 (ab11408; Abcam) and anti-human IFN-β (ab6979; Abcam) were added to GILZ RNAi cell culture or RNAi controls 3 h before IBDV infection.

    Techniques: Expressing

    GILZ mediates the inhibitory effect of VP4 on TNF-α or SeV-induced activation of IFN-α, IFN-β, and NF-κB promoters. (A and B) Effects of GILZ RNAi on the expression of endogenous GILZ. HEK293T cells (2.0 × 10 5 )

    Journal: Journal of Virology

    Article Title: Critical Roles of Glucocorticoid-Induced Leucine Zipper in Infectious Bursal Disease Virus (IBDV)-Induced Suppression of Type I Interferon Expression and Enhancement of IBDV Growth in Host Cells via Interaction with VP4

    doi: 10.1128/JVI.02421-12

    Figure Lengend Snippet: GILZ mediates the inhibitory effect of VP4 on TNF-α or SeV-induced activation of IFN-α, IFN-β, and NF-κB promoters. (A and B) Effects of GILZ RNAi on the expression of endogenous GILZ. HEK293T cells (2.0 × 10 5 )

    Article Snippet: To neutralize interferon's activity, 2 μg of anti-human IFN-α1 (ab11408; Abcam) and anti-human IFN-β (ab6979; Abcam) were added to GILZ RNAi cell culture or RNAi controls 3 h before IBDV infection.

    Techniques: Activation Assay, Expressing

    Loss of Dicer triggers the interferon response a) Western blot analysis of cell extracts isolated from normal and Dicer knockdown HEK293 cells, cultured for 1 or 2 weeks, using antibodies specific to Dicer, TLR3, PKR1 and β-actin. Anti-TLR3 antibody detects two bands and according to size markers, the upper band is likely to correspond to modified protein. Quantitation is shown in Supplementary Figure 7 . See Supplementary Fig. 9 for uncropped blot images. b) Western blot analysis of cell extracts isolated from normal and Dicer knockdown HEK293 cells, cultured for 10 days using antibodies specific to IFN-β, OAS1 and tubulin. See Supplementary Fig. 9 for uncropped blot images. c) Western blot analysis as in (b) using antibodies specific to ADAR1 and tubulin. See Supplementary Fig. 9 for uncropped blot images. d) Western blot analysis of cell extracts isolated from normal and siRNA-directed Dicer knockdown HEK293 cells using antibodies specific to OAS1, TLR3 and tubulin. See Supplementary Fig. 9 for uncropped blot images. e) Western blot analysis of cell extracts isolated from normal and Drosha knockdown HEK293 cells using antibodies specific to Drosha, TLR3 and tubulin. See Supplementary Fig. 9 for uncropped blot images. f) Flow cytometry based quantitation of the % of cell population undergoing apoptosis in normal and Dicer, Drosha or PKR knockdown cells. % of cell population values are based on average values ± s.e.m. from three independent biological experiments. All experiments in the figure were independently repeated 3 times.

    Journal: Nature structural & molecular biology

    Article Title: HUMAN NUCLEAR DICER RESTRICTS THE DELETERIOUS ACCUMULATION OF ENDOGENOUS DOUBLE STRAND RNA

    doi: 10.1038/nsmb.2827

    Figure Lengend Snippet: Loss of Dicer triggers the interferon response a) Western blot analysis of cell extracts isolated from normal and Dicer knockdown HEK293 cells, cultured for 1 or 2 weeks, using antibodies specific to Dicer, TLR3, PKR1 and β-actin. Anti-TLR3 antibody detects two bands and according to size markers, the upper band is likely to correspond to modified protein. Quantitation is shown in Supplementary Figure 7 . See Supplementary Fig. 9 for uncropped blot images. b) Western blot analysis of cell extracts isolated from normal and Dicer knockdown HEK293 cells, cultured for 10 days using antibodies specific to IFN-β, OAS1 and tubulin. See Supplementary Fig. 9 for uncropped blot images. c) Western blot analysis as in (b) using antibodies specific to ADAR1 and tubulin. See Supplementary Fig. 9 for uncropped blot images. d) Western blot analysis of cell extracts isolated from normal and siRNA-directed Dicer knockdown HEK293 cells using antibodies specific to OAS1, TLR3 and tubulin. See Supplementary Fig. 9 for uncropped blot images. e) Western blot analysis of cell extracts isolated from normal and Drosha knockdown HEK293 cells using antibodies specific to Drosha, TLR3 and tubulin. See Supplementary Fig. 9 for uncropped blot images. f) Flow cytometry based quantitation of the % of cell population undergoing apoptosis in normal and Dicer, Drosha or PKR knockdown cells. % of cell population values are based on average values ± s.e.m. from three independent biological experiments. All experiments in the figure were independently repeated 3 times.

    Article Snippet: Western blot experiments were performed according to standard protocols using the following antibodies: anti-Pol II 8WG16 (Abcam-ab24758), anti-Pol II N20 (Santa Cruz Biotechnology-sc-899), anti-Dicer 13D6 (Abcam- ab14601), anti-β actin (Sigma-A3853), anti-Spt5 (Millipore-ABE443), anti-TLR3 (Millipore-06-008), anti-PKR1 (Millipore-07-151), anti-IFN-β (Abcam-ab6979), anti-OAS1 (Abcam-ab82666), anti-ADAR1 (Abcam-ab88574), anti-Drosha (Abcamab12286), anti-tubulin (Sigma-T5168), anti-Grp75 (Abcam-ab2799).

    Techniques: Western Blot, Isolation, Cell Culture, Modification, Protein Quantitation, Flow Cytometry, Cytometry, Quantitation Assay