l phenylalanine  (Millipore)


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    Name:
    L Phenylalanine
    Description:

    Catalog Number:
    p2126
    Price:
    None
    Applications:
    L-Phenylalanine has been used in Fluo-4 Ca2+-assay in a study to demonstrate that G-protein coupled receptor 139 reference surrogate agonists 1a and 7c and L-phenylalanine share a common binding site. It has also been used to spike blood samples to study the effect of dried blood spots on newborn screening.
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    Structured Review

    Millipore l phenylalanine
    L Phenylalanine

    https://www.bioz.com/result/l phenylalanine/product/Millipore
    Average 90 stars, based on 32 article reviews
    Price from $9.99 to $1999.99
    l phenylalanine - by Bioz Stars, 2020-01
    90/100 stars

    Images

    1) Product Images from "N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor"

    Article Title: N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.00574

    Effects of the NOX-inhibitor diphenyleneiodonium (DPI) on  N -formyl peptide receptor (FPR)-mediated reactive oxygen species (ROS) production and FPRs-induced modulation of gp91 phox  and p67 phox  expression in BJ cells.  (A)  BJ cells were plated in a 96-well plate and pretreated for 1 h at 37°C with 10 µM of the NOX-inhibitor DPI. At the end of incubation, cells were treated with  N -formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm peptide, uPAR 84–95  peptide, or TGF-β. Results are expressed as percentage of increase of mean fluorescence intensity of stimulated DCHF-DA-loaded cells in respect to unstimulated DCHF-DA-loaded cells (considered as 100%). Values are the mean ± SEM of three experiments performed in triplicate. ** p
    Figure Legend Snippet: Effects of the NOX-inhibitor diphenyleneiodonium (DPI) on N -formyl peptide receptor (FPR)-mediated reactive oxygen species (ROS) production and FPRs-induced modulation of gp91 phox and p67 phox expression in BJ cells. (A) BJ cells were plated in a 96-well plate and pretreated for 1 h at 37°C with 10 µM of the NOX-inhibitor DPI. At the end of incubation, cells were treated with N -formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm peptide, uPAR 84–95 peptide, or TGF-β. Results are expressed as percentage of increase of mean fluorescence intensity of stimulated DCHF-DA-loaded cells in respect to unstimulated DCHF-DA-loaded cells (considered as 100%). Values are the mean ± SEM of three experiments performed in triplicate. ** p

    Techniques Used: Expressing, Incubation, Fluorescence

    Effects of N -Formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm peptide, and uPAR 84–95 on reactive oxygen species (ROS) production in BJ cells. Cells were plated in a 96-well plate and treated with DCHF-DA. At the end of incubation, cells were treated with medium alone (black columns), fMLF (dark gray columns), WKYMVm peptide (white columns), and uPAR 84–95 peptide (light gray columns). ROS release was measured as dichlorofluorescein (DCF) fluorescence intensity at 5, 15, 30, and 60 min (A–D) . Results are expressed as mean fluorescence intensity of DCHF-DA-loaded cells. DCHF-DA-loaded unstimulated cells and TGF-β stimulated cells were examined in parallel, as controls, and are shown in insets. Values are the mean ± SEM of three experiments performed in triplicate. * p
    Figure Legend Snippet: Effects of N -Formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm peptide, and uPAR 84–95 on reactive oxygen species (ROS) production in BJ cells. Cells were plated in a 96-well plate and treated with DCHF-DA. At the end of incubation, cells were treated with medium alone (black columns), fMLF (dark gray columns), WKYMVm peptide (white columns), and uPAR 84–95 peptide (light gray columns). ROS release was measured as dichlorofluorescein (DCF) fluorescence intensity at 5, 15, 30, and 60 min (A–D) . Results are expressed as mean fluorescence intensity of DCHF-DA-loaded cells. DCHF-DA-loaded unstimulated cells and TGF-β stimulated cells were examined in parallel, as controls, and are shown in insets. Values are the mean ± SEM of three experiments performed in triplicate. * p

    Techniques Used: Incubation, Fluorescence

    Effects of the inhibition of the cross-talk between FPRs-uPAR-Integrins on reactive oxygen species (ROS) induction in BJ cells.  (A)  Western blot analysis of uPAR expression in H460 cell line as a positive control (lane 1), BJ cells at first passage (lane 2) and fifth passage (lane 3) in culture with the R4 anti-uPAR mAb and with an anti-β-actin Ab, as a loading control.  (B)  BJ cells were plated in a 96-well plate and treated with DCHF-DA. At the end of incubation, cells were treated with medium alone,  N -formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm or TGF-β in the absence (black columns) or in the presence of nonimmune immunoglobulins (medium gray columns), IB antibody (dark gray columns), scrambled control peptide (Scp) (white columns), and P25 peptide (light gray columns). ROS release was measured as dichlorofluorescein (DCF) fluorescence at 30 min. Results are expressed as a percentage of increase of mean fluorescence intensity of stimulated DCHF-DA-loaded cells in respect to unstimulated DCHF-DA-loaded cells (considered as 100%). Values are the mean ± SEM of three experiments performed in triplicate. * p
    Figure Legend Snippet: Effects of the inhibition of the cross-talk between FPRs-uPAR-Integrins on reactive oxygen species (ROS) induction in BJ cells. (A) Western blot analysis of uPAR expression in H460 cell line as a positive control (lane 1), BJ cells at first passage (lane 2) and fifth passage (lane 3) in culture with the R4 anti-uPAR mAb and with an anti-β-actin Ab, as a loading control. (B) BJ cells were plated in a 96-well plate and treated with DCHF-DA. At the end of incubation, cells were treated with medium alone, N -formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm or TGF-β in the absence (black columns) or in the presence of nonimmune immunoglobulins (medium gray columns), IB antibody (dark gray columns), scrambled control peptide (Scp) (white columns), and P25 peptide (light gray columns). ROS release was measured as dichlorofluorescein (DCF) fluorescence at 30 min. Results are expressed as a percentage of increase of mean fluorescence intensity of stimulated DCHF-DA-loaded cells in respect to unstimulated DCHF-DA-loaded cells (considered as 100%). Values are the mean ± SEM of three experiments performed in triplicate. * p

    Techniques Used: Inhibition, Western Blot, Expressing, Positive Control, Incubation, Fluorescence

    FPRs-mediated Rac1 and ERK activation in BJ cells. (A) BJ cells, treated with medium alone (-), N -formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm peptide, or uPAR 84–95 peptide, were lysed and subjected to Rac1 activity assay using PAK-PBD-glutathione sepharose beads. Immunoprecipitates and the corresponding total lysates, as an input control, were subjected to Western blot analysis with an anti-Rac1-specific Ab and with an anti-β-actin Ab, as a loading control. (B) BJ cells, treated with medium alone, fMLF, WKYMV peptide, or uPAR 84–95 peptide were lysed and subjected to Western blot analysis with an anti phospho-ERK 1/2 (p-ERK)-specific Ab and then with anti ERK-2 and anti-β-actin Abs, as a loading control. (C) BJ cells were plated in a 96-well plate and treated with DCHF-DA. At the end of incubation, cells were treated with fMLF, WKYMVm peptide, and uPAR 84–95 peptide in the absence (white columns) or in the presence of NSC23766 (black columns) or PD98059 (gray columns). ROS release was measured as dichlorofluorescein (DCF) fluorescence intensity at 5 min. Results are expressed as a percentage of increase of mean fluorescence intensity of stimulated DCHF-DA-loaded cells in respect to unstimulated DCHF-DA-loaded cells (considered as 100%). Values are the mean ± SEM of three experiments performed in triplicate. * p
    Figure Legend Snippet: FPRs-mediated Rac1 and ERK activation in BJ cells. (A) BJ cells, treated with medium alone (-), N -formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm peptide, or uPAR 84–95 peptide, were lysed and subjected to Rac1 activity assay using PAK-PBD-glutathione sepharose beads. Immunoprecipitates and the corresponding total lysates, as an input control, were subjected to Western blot analysis with an anti-Rac1-specific Ab and with an anti-β-actin Ab, as a loading control. (B) BJ cells, treated with medium alone, fMLF, WKYMV peptide, or uPAR 84–95 peptide were lysed and subjected to Western blot analysis with an anti phospho-ERK 1/2 (p-ERK)-specific Ab and then with anti ERK-2 and anti-β-actin Abs, as a loading control. (C) BJ cells were plated in a 96-well plate and treated with DCHF-DA. At the end of incubation, cells were treated with fMLF, WKYMVm peptide, and uPAR 84–95 peptide in the absence (white columns) or in the presence of NSC23766 (black columns) or PD98059 (gray columns). ROS release was measured as dichlorofluorescein (DCF) fluorescence intensity at 5 min. Results are expressed as a percentage of increase of mean fluorescence intensity of stimulated DCHF-DA-loaded cells in respect to unstimulated DCHF-DA-loaded cells (considered as 100%). Values are the mean ± SEM of three experiments performed in triplicate. * p

    Techniques Used: Activation Assay, Activity Assay, Western Blot, Incubation, Fluorescence

    FPRs-mediated Rac1 GTP-p67 phox  interaction in BJ cells.  (A)  BJ cells lysates were treated with GDP and GTPγS (Upstate) to generate Rac1-GDP and Rac1-GTP, respectively. Rac1-GDP and Rac1-GTP containing lysates were precipitated using the p21-binding domain (PBD) of PAK1, bound to agarose beads. Eluted samples and the corresponding total lysates were subjected to Western blot analysis with a polyclonal anti-p67 phox  antibody and with anti-Rac1 antibody as a loading control, respectively.  (B)  BJ cells, after incubation with medium alone (-), N -formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm peptide, for 5 min, or uPAR 84–95  (10 −8 M) for 30 min at 37°C in a humidified (5% CO 2 ) incubator, were lysed and subjected to Rac1 activity assay. Active Rac1 (Rac1-GTP) was precipitated from cell lysates using the PBD of PAK1, bound to agarose beads. Eluted samples and the corresponding total lysates were subjected to Western blot analysis with a polyclonal anti-p67 phox  antibody and with anti-Rac1 antibody as a loading control, respectively.
    Figure Legend Snippet: FPRs-mediated Rac1 GTP-p67 phox interaction in BJ cells. (A) BJ cells lysates were treated with GDP and GTPγS (Upstate) to generate Rac1-GDP and Rac1-GTP, respectively. Rac1-GDP and Rac1-GTP containing lysates were precipitated using the p21-binding domain (PBD) of PAK1, bound to agarose beads. Eluted samples and the corresponding total lysates were subjected to Western blot analysis with a polyclonal anti-p67 phox antibody and with anti-Rac1 antibody as a loading control, respectively. (B) BJ cells, after incubation with medium alone (-), N -formyl- l -methionyl- l -leucyl- l -phenylalanine (fMLF), WKYMVm peptide, for 5 min, or uPAR 84–95 (10 −8 M) for 30 min at 37°C in a humidified (5% CO 2 ) incubator, were lysed and subjected to Rac1 activity assay. Active Rac1 (Rac1-GTP) was precipitated from cell lysates using the PBD of PAK1, bound to agarose beads. Eluted samples and the corresponding total lysates were subjected to Western blot analysis with a polyclonal anti-p67 phox antibody and with anti-Rac1 antibody as a loading control, respectively.

    Techniques Used: Binding Assay, Western Blot, Incubation, Activity Assay

    Related Articles

    Negative Control:

    Article Title: Heteroconium chaetospira Induces Resistance to Clubroot via Upregulation of Host Genes Involved in Jasmonic Acid, Ethylene, and Auxin Biosynthesis
    Article Snippet: There were 6 treatments for this analysis: negative control (formulation blank, no Pb), pathogen control (formulation blank + Pb), BC2HB1 alone at 1.25% and 2.5%, and BC2HB1 at 1.25% and 2.5% plus Pb, respectively. .. An assay mixture consisted of 1.5 ml borate buffer (150 μM), 1.0 ml deionised water, 1 ml L-phenylalanine (Sigma–Aldrich Canada, Oakville, ON) solution (10 μM) and 0.5 ml the supernatant.

    Radioactivity:

    Article Title: Autoubiquitination of the 26S Proteasome on Rpn13 Regulates Breakdown of Ubiquitin Conjugates
    Article Snippet: To measure the degradation rate of cellular proteins, HEK293 cells were first labeled for 24 h with tritium-labeled phenylalanine (Phe L-[3,4,5 3 H], American Radiolabeled Chemicals, Inc. ART0614, stock 1 mCi/ml in 0.01 M HCl, final 5 μCi/ml) and then chased for 1 h with complete media containing 2 mM cold phenylalanine (Sigma, P5482-25G) to allow turnover of short-lived proteins. .. 200 μl supernatant after TCA precipitation was mixed with 3 ml Ultima Gold Scintillation Fluid (PerkinElmer, 6013327) and TCA-soluble radioactivity was measured with a PerkinElmer Tri-Carb 2910TR Liquid Scintillation Analyzer equipped with QuantaSmart™ software.

    TCA Precipitation:

    Article Title: Rapid induction of p62 and GABARAPL1 upon proteasome inhibition promotes survival before autophagy activation
    Article Snippet: Measurement of cellular protein degradation To measure the degradation rate of long-lived proteins, WT or sh-p62 SH-SY5Y cells were first labeled for 24 h with [3 H]phenylalanine (Phe l -[3,4,5 3 H], ART0614, stock 1 mCi/ml in 0.01 M HCl, final 2 µCi/ml; American Radiolabeled Chemicals) and then chased for 2 h with complete medium containing 2 mM nonradioactive phenylalanine (P5482-25G; Sigma-Aldrich) to allow degradation of short-lived radioactive proteins. .. After TCA precipitation, 200 µl supernatant was mixed with 3 ml Ultima gold scintillation fluid (6013327; PerkinElmer) and measured with a PerkinElmer Tri-Carb 2910TR liquid scintillation analyzer.

    Article Title: Autoubiquitination of the 26S Proteasome on Rpn13 Regulates Breakdown of Ubiquitin Conjugates
    Article Snippet: To measure the degradation rate of cellular proteins, HEK293 cells were first labeled for 24 h with tritium-labeled phenylalanine (Phe L-[3,4,5 3 H], American Radiolabeled Chemicals, Inc. ART0614, stock 1 mCi/ml in 0.01 M HCl, final 5 μCi/ml) and then chased for 1 h with complete media containing 2 mM cold phenylalanine (Sigma, P5482-25G) to allow turnover of short-lived proteins. .. 200 μl supernatant after TCA precipitation was mixed with 3 ml Ultima Gold Scintillation Fluid (PerkinElmer, 6013327) and TCA-soluble radioactivity was measured with a PerkinElmer Tri-Carb 2910TR Liquid Scintillation Analyzer equipped with QuantaSmart™ software.

    Article Title: Rapid induction of p62 and GABARAPL1 upon proteasome inhibition promotes survival before autophagy activation
    Article Snippet: To measure the degradation rate of long-lived proteins, WT or sh-p62 SH-SY5Y cells were first labeled for 24 h with [3 H]phenylalanine (Phe l -[3,4,5 3 H], ART0614, stock 1 mCi/ml in 0.01 M HCl, final 2 µCi/ml; American Radiolabeled Chemicals) and then chased for 2 h with complete medium containing 2 mM nonradioactive phenylalanine (P5482-25G; Sigma-Aldrich) to allow degradation of short-lived radioactive proteins. .. After TCA precipitation, 200 µl supernatant was mixed with 3 ml Ultima gold scintillation fluid (6013327; PerkinElmer) and measured with a PerkinElmer Tri-Carb 2910TR liquid scintillation analyzer.

    Isolation:

    Article Title: Cytoplasmic flows in starfish oocytes are fully determined by cortical contractions
    Article Snippet: Oocytes are isolated from a biopsy of the ovaries that is obtained from the dorsal side of a starfish arm using a surgical biopsy puncher. .. The biopsy is put into calcium-free sea water at pH 8 with 50 mM L-Phenylalanine (Sigma) added for 15 min to 20 min.

    Flow Cytometry:

    Article Title: Glutamine and Leucine Provide Enhanced Protective Immunity Against Mucosal Infection with Herpes Simplex Virus Type 1
    Article Snippet: Reagents and antibodies L-Gln (G-3126), L-Leu (L8912), L-glutamic acid (Glu) (G8415), L-phenylalanine (Phe) (P5482), L-lysine (Lys) (L5501), L-asparagine (Asn) (A0884), and L-Arg were purchased from Sigma. .. The mAbs used for the flow cytometric analysis and other experiments were prepared from eBioscience (San Diego, CA) or BD Bioscience (San Diego, CA), and included: FITC-anti-mouse CD3e (145-2C11), CD4 (Rm4-5), CD8 (53-6.7) CD44 (IM7), CD62L (MEL-14), CD69 (H1.2F3), CD80 (16-10A1), phycoerythrin (PE) conjugated anti-mouse- CD4 (RM4-5) or CD8 (53-6.7), CD19 (MB19-1), CD154-(CD40L) (MR1), IFN-γ (XMG1.2), granzyme B (16G6), peridinin chlorophyll protein complex (PerCP)-conjugated anti-mouse IFN-γ, allophycocyanin (APC)-conjugated anti-mouse TNF-α, biotin-conjugated anti-mouse pan-NK cells (CD49b) [DX5], Streptavidin-APC.

    Cell Culture:

    Article Title: Enhanced Cytotoxicity from Deoxyguanosine-Enriched T-oligo in Prostate Cancer Cells
    Article Snippet: Paragraph title: Cell culture ... The LNCaP cell line was maintained in 10% fetal bovine serum (Hyclone); RPMI1640 medium (Gibco); 2 mM l -glutamine (Gibco); 200 U penicillin/mL; 200 μg streptomycin/mL (Gibco). pZ-HPV-7 cells were maintained in MCDB-153 medium supplemented with 0.05 mg/mL l -histidine, 0.099 mg/mL l -isoleucine, 0.014 mg/mL l -methionine, 0.010 mg/mL l -tryptophan, 0.014 mg/mL l -tyrosine, 0.015 mg/mL l -phenylalanine, 5 μg/mL bovine insulin, and 50 ng/mL prostaglandin E1 all purchased from Sigma-Aldrich.

    Purification:

    Article Title: Engineering of near infrared fluorescent proteinoid-poly(L-lactic acid) particles for in vivo colon cancer detection
    Article Snippet: .. Materials The following analytical-grade chemicals were purchased from commercial sources and were used without further purification: L-glutamic acid (E), L-phenylalanine (F), indiocyaninegreen (ICG), human serum albumin (HSA), bovine plasma fibrinogen, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), Matrigel, Triton-x-100 and monoclonal anti-CEA antibodies (T86-66) from Sigma (Rehovot, Israel); Poly(L-lactic acid) MW 2,000 Da from Polysciences (Warrington, PA, USA); N -hydroxysulfosuccinimide (Sulfo-NHS) and 2-morpholino ethanesulfonic acid (MES, pH 6) from Thermo Fisher Scientific (Rockford, IL, USA); Phosphate Buffered Saline (PBS), Minimum Essential Medium (MEM) eagle, McCoy’s 5A medium and Dulbecco’s modification of eagle’s medium (DMEM), Fetal Bovine Serum (FBS), glutamine, penicillin, streptomycin and mycoplasma detection kit from Biological Industries (Bet Haemek, Israel); cell cytotoxicity assay kit (LDH) from Roche (Switzerland); LS174t, SW480 and HT29 cell lines from American Type Culture Collection (ATCC); donkey anti-rabbit IgG from Jackson ImmunoResearch Laboratories (West Grove, PA, USA); water was purified by passing deionized water through an Elgastat Spectrum reverse osmosis system (Elga Ltd., High Wycombe, UK). ..

    Labeling:

    Article Title: Rapid induction of p62 and GABARAPL1 upon proteasome inhibition promotes survival before autophagy activation
    Article Snippet: .. Measurement of cellular protein degradation To measure the degradation rate of long-lived proteins, WT or sh-p62 SH-SY5Y cells were first labeled for 24 h with [3 H]phenylalanine (Phe l -[3,4,5 3 H], ART0614, stock 1 mCi/ml in 0.01 M HCl, final 2 µCi/ml; American Radiolabeled Chemicals) and then chased for 2 h with complete medium containing 2 mM nonradioactive phenylalanine (P5482-25G; Sigma-Aldrich) to allow degradation of short-lived radioactive proteins. .. Cells were then incubated in chase medium containing 1 µM BTZ.

    Article Title: Autoubiquitination of the 26S Proteasome on Rpn13 Regulates Breakdown of Ubiquitin Conjugates
    Article Snippet: .. To measure the degradation rate of cellular proteins, HEK293 cells were first labeled for 24 h with tritium-labeled phenylalanine (Phe L-[3,4,5 3 H], American Radiolabeled Chemicals, Inc. ART0614, stock 1 mCi/ml in 0.01 M HCl, final 5 μCi/ml) and then chased for 1 h with complete media containing 2 mM cold phenylalanine (Sigma, P5482-25G) to allow turnover of short-lived proteins. .. Cells were then allowed to be incubated in media containing both 2 mM cold Phe and various concentrations of BTZ.

    Article Title: Rapid induction of p62 and GABARAPL1 upon proteasome inhibition promotes survival before autophagy activation
    Article Snippet: .. To measure the degradation rate of long-lived proteins, WT or sh-p62 SH-SY5Y cells were first labeled for 24 h with [3 H]phenylalanine (Phe l -[3,4,5 3 H], ART0614, stock 1 mCi/ml in 0.01 M HCl, final 2 µCi/ml; American Radiolabeled Chemicals) and then chased for 2 h with complete medium containing 2 mM nonradioactive phenylalanine (P5482-25G; Sigma-Aldrich) to allow degradation of short-lived radioactive proteins. .. Cells were then incubated in chase medium containing 1 µM BTZ.

    Concentration Assay:

    Article Title: The GPR139 reference agonists 1a and 7c, and tryptophan and phenylalanine share a common binding site
    Article Snippet: .. DMSO was confirmed not to have any activity by itself at this concentration . l - Trp (T0254) and l -Phe (P2126) were obtained from Sigma-Aldrich and dissolved in buffer. .. Fluo-4 Ca2+ -assay The Ca2+ measurements were preformed using the Fluo-4 NW Calcium Assay Kit (Invitrogen, Molecular Probes, F36206) as previously described .

    Incubation:

    Article Title: Rapid induction of p62 and GABARAPL1 upon proteasome inhibition promotes survival before autophagy activation
    Article Snippet: Measurement of cellular protein degradation To measure the degradation rate of long-lived proteins, WT or sh-p62 SH-SY5Y cells were first labeled for 24 h with [3 H]phenylalanine (Phe l -[3,4,5 3 H], ART0614, stock 1 mCi/ml in 0.01 M HCl, final 2 µCi/ml; American Radiolabeled Chemicals) and then chased for 2 h with complete medium containing 2 mM nonradioactive phenylalanine (P5482-25G; Sigma-Aldrich) to allow degradation of short-lived radioactive proteins. .. Cells were then incubated in chase medium containing 1 µM BTZ.

    Article Title: Heteroconium chaetospira Induces Resistance to Clubroot via Upregulation of Host Genes Involved in Jasmonic Acid, Ethylene, and Auxin Biosynthesis
    Article Snippet: An assay mixture consisted of 1.5 ml borate buffer (150 μM), 1.0 ml deionised water, 1 ml L-phenylalanine (Sigma–Aldrich Canada, Oakville, ON) solution (10 μM) and 0.5 ml the supernatant. .. After incubation at 38°C for 2 h, 50 μl of 5N HCL was added to the mixture to stop the reaction.

    Article Title: Effect of Cadmium and Copper Exposure on Growth, Secondary Metabolites and Antioxidant Activity in the Medicinal Plant Sambung Nyawa (Gynura procumbens (Lour.) Merr)
    Article Snippet: .. Enzyme extract (10 μL) was incubated at 40 °C with 12.1 mM l -phenylalanine (90 μL, Sigma; St Louis, MS, USA) that was prepared in 50 mM Tris-HCl, (pH 8.5). ..

    Article Title: Abscisic Acid Induced Changes in Production of Primary and Secondary Metabolites, Photosynthetic Capacity, Antioxidant Capability, Antioxidant Enzymes and Lipoxygenase Inhibitory Activity of Orthosiphon stamineus Benth.
    Article Snippet: .. Enzyme extract (10 µL) was incubated at 40 °C with 12.1 mM L-phenylalanine (90 µL, Sigma) that was prepared in 50 mM Tris-HCl, (pH 8.5). ..

    Article Title: Autoubiquitination of the 26S Proteasome on Rpn13 Regulates Breakdown of Ubiquitin Conjugates
    Article Snippet: To measure the degradation rate of cellular proteins, HEK293 cells were first labeled for 24 h with tritium-labeled phenylalanine (Phe L-[3,4,5 3 H], American Radiolabeled Chemicals, Inc. ART0614, stock 1 mCi/ml in 0.01 M HCl, final 5 μCi/ml) and then chased for 1 h with complete media containing 2 mM cold phenylalanine (Sigma, P5482-25G) to allow turnover of short-lived proteins. .. Cells were then allowed to be incubated in media containing both 2 mM cold Phe and various concentrations of BTZ.

    Article Title: Rapid induction of p62 and GABARAPL1 upon proteasome inhibition promotes survival before autophagy activation
    Article Snippet: To measure the degradation rate of long-lived proteins, WT or sh-p62 SH-SY5Y cells were first labeled for 24 h with [3 H]phenylalanine (Phe l -[3,4,5 3 H], ART0614, stock 1 mCi/ml in 0.01 M HCl, final 2 µCi/ml; American Radiolabeled Chemicals) and then chased for 2 h with complete medium containing 2 mM nonradioactive phenylalanine (P5482-25G; Sigma-Aldrich) to allow degradation of short-lived radioactive proteins. .. Cells were then incubated in chase medium containing 1 µM BTZ.

    other:

    Article Title: A post-ingestive amino acid sensor promotes food consumption in Drosophila
    Article Snippet: Chemicals Sucrose (S7903), agar (A1296), L-alanine (A7627), L-argine (A5131), L-asparagine (A0884), L-aspartate (A8949), L-cysteine (C1276), L-glutamate (G1251), L-glutamine (G3126), L-glycine (G7126), L-histidine (H8000), L-isoleucine (I2752), L-leucine (L8912), L- methionine (M9625), L-phenylalanine (P2126), L-proline (P0380), L-serine (S4500), L-threonine (T8625), L-tryptophan (T0254), L-tyrosine (T3754), L-valine (V0500), L-lysine (L5626), D-alanine (A7377), D-aspartate (V900627), D-glutamate (G1001), denatomium (D5765), and protein A (P6031) were purchased from Sigma-Aldrich.

    Article Title: A Highly Sensitive Nonenzymatic Glucose Biosensor Based on the Regulatory Effect of Glucose on Electrochemical Behaviors of Colloidal Silver Nanoparticles on MoS2†
    Article Snippet: Reagents and Materials Silver nitrite ( > 99% AgNO3 ), sodium borohydride (99% NaBH4 ), glucose, glycine, urea, l -phenylalanine, l -lactic acid and l -tyrosine were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Article Title: Autophagy induction by tetrahydrobiopterin deficiency
    Article Snippet: L-DOPA (D9628), 5-hydroxytryptophan (H9772), carbidopa (C1335), ascorbic acid (A4544), N-acetyl-L-cysteine (A8199), L-tyrosine (T3754), L-leucine (L8912), L-phenylalanine (P5482), L-glutamine (G8540) and anti-α-tubulin antibody (T5168) were purchased from Sigma.

    Activity Assay:

    Article Title: Heteroconium chaetospira Induces Resistance to Clubroot via Upregulation of Host Genes Involved in Jasmonic Acid, Ethylene, and Auxin Biosynthesis
    Article Snippet: Paragraph title: Activity of phenylalanine ammonia lyases (PAL) ... An assay mixture consisted of 1.5 ml borate buffer (150 μM), 1.0 ml deionised water, 1 ml L-phenylalanine (Sigma–Aldrich Canada, Oakville, ON) solution (10 μM) and 0.5 ml the supernatant.

    Article Title: The GPR139 reference agonists 1a and 7c, and tryptophan and phenylalanine share a common binding site
    Article Snippet: .. DMSO was confirmed not to have any activity by itself at this concentration . l - Trp (T0254) and l -Phe (P2126) were obtained from Sigma-Aldrich and dissolved in buffer. .. Fluo-4 Ca2+ -assay The Ca2+ measurements were preformed using the Fluo-4 NW Calcium Assay Kit (Invitrogen, Molecular Probes, F36206) as previously described .

    Article Title: Effect of Cadmium and Copper Exposure on Growth, Secondary Metabolites and Antioxidant Activity in the Medicinal Plant Sambung Nyawa (Gynura procumbens (Lour.) Merr)
    Article Snippet: Paragraph title: 3.8. Phenylalanine Ammonia-Lyase (PAL) Activity ... Enzyme extract (10 μL) was incubated at 40 °C with 12.1 mM l -phenylalanine (90 μL, Sigma; St Louis, MS, USA) that was prepared in 50 mM Tris-HCl, (pH 8.5).

    Article Title: Abscisic Acid Induced Changes in Production of Primary and Secondary Metabolites, Photosynthetic Capacity, Antioxidant Capability, Antioxidant Enzymes and Lipoxygenase Inhibitory Activity of Orthosiphon stamineus Benth.
    Article Snippet: Paragraph title: 3.8. Phenylalanine Ammonia-Lyase (PAL) Activity ... Enzyme extract (10 µL) was incubated at 40 °C with 12.1 mM L-phenylalanine (90 µL, Sigma) that was prepared in 50 mM Tris-HCl, (pH 8.5).

    Article Title: Biochemical Evaluation of Phenylalanine Ammonia Lyase from Endemic Plant Cyathobasis fruticulosa (Bunge) Aellen. for the Dietary Treatment of Phenylketonuria
    Article Snippet: Paragraph title: PAL activity assay and biochemical property analysis ... The enzyme reaction mixture contained 100 mM Tris- -HCl, 40 mM l -phenylalanine, ≥98% (Sigma-Aldrich), and an aliquot of the enzyme in a total volume of 1 mL at pH=8.8.

    Spectrophotometry:

    Article Title: Heteroconium chaetospira Induces Resistance to Clubroot via Upregulation of Host Genes Involved in Jasmonic Acid, Ethylene, and Auxin Biosynthesis
    Article Snippet: An assay mixture consisted of 1.5 ml borate buffer (150 μM), 1.0 ml deionised water, 1 ml L-phenylalanine (Sigma–Aldrich Canada, Oakville, ON) solution (10 μM) and 0.5 ml the supernatant. .. The PAL activity was quantified by measuring the total amount of t-cinnamic acid formed in each sample using a SPECTRAmax Plus spectrophotometer (Molecular Devices, Sunnyvale, CA) at 290 nm relative to a reference PAL activity in controls without the addition of L- phenylalanine , .

    Article Title: Biochemical Evaluation of Phenylalanine Ammonia Lyase from Endemic Plant Cyathobasis fruticulosa (Bunge) Aellen. for the Dietary Treatment of Phenylketonuria
    Article Snippet: PAL activity was assayed by measuring the trans -cinnamic acid formation at 290 nm using a UV-1800 UV-Vis spectrophotometer (Shimadzu, Kyoto, Japan) and calculated using a standard trans -cinnamic acid, ≥99% (Sigma-Aldrich) curve. .. The enzyme reaction mixture contained 100 mM Tris- -HCl, 40 mM l -phenylalanine, ≥98% (Sigma-Aldrich), and an aliquot of the enzyme in a total volume of 1 mL at pH=8.8.

    Mass Spectrometry:

    Article Title: Effect of Cadmium and Copper Exposure on Growth, Secondary Metabolites and Antioxidant Activity in the Medicinal Plant Sambung Nyawa (Gynura procumbens (Lour.) Merr)
    Article Snippet: .. Enzyme extract (10 μL) was incubated at 40 °C with 12.1 mM l -phenylalanine (90 μL, Sigma; St Louis, MS, USA) that was prepared in 50 mM Tris-HCl, (pH 8.5). ..

    Modification:

    Article Title: Engineering of near infrared fluorescent proteinoid-poly(L-lactic acid) particles for in vivo colon cancer detection
    Article Snippet: .. Materials The following analytical-grade chemicals were purchased from commercial sources and were used without further purification: L-glutamic acid (E), L-phenylalanine (F), indiocyaninegreen (ICG), human serum albumin (HSA), bovine plasma fibrinogen, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), Matrigel, Triton-x-100 and monoclonal anti-CEA antibodies (T86-66) from Sigma (Rehovot, Israel); Poly(L-lactic acid) MW 2,000 Da from Polysciences (Warrington, PA, USA); N -hydroxysulfosuccinimide (Sulfo-NHS) and 2-morpholino ethanesulfonic acid (MES, pH 6) from Thermo Fisher Scientific (Rockford, IL, USA); Phosphate Buffered Saline (PBS), Minimum Essential Medium (MEM) eagle, McCoy’s 5A medium and Dulbecco’s modification of eagle’s medium (DMEM), Fetal Bovine Serum (FBS), glutamine, penicillin, streptomycin and mycoplasma detection kit from Biological Industries (Bet Haemek, Israel); cell cytotoxicity assay kit (LDH) from Roche (Switzerland); LS174t, SW480 and HT29 cell lines from American Type Culture Collection (ATCC); donkey anti-rabbit IgG from Jackson ImmunoResearch Laboratories (West Grove, PA, USA); water was purified by passing deionized water through an Elgastat Spectrum reverse osmosis system (Elga Ltd., High Wycombe, UK). ..

    Article Title: Enhanced Cytotoxicity from Deoxyguanosine-Enriched T-oligo in Prostate Cancer Cells
    Article Snippet: DU145, PC-3, and LN-4 cells were maintained in 10% fetal bovine serum purchased from Hyclone; Dulbecco's modified Eagle's medium (DMEM) from Mediatech; 2 mM l -Glutamine from Gibco; 200 U/mL penicillin; 200 μg/mL streptomycin (Gibco). .. The LNCaP cell line was maintained in 10% fetal bovine serum (Hyclone); RPMI1640 medium (Gibco); 2 mM l -glutamine (Gibco); 200 U penicillin/mL; 200 μg streptomycin/mL (Gibco). pZ-HPV-7 cells were maintained in MCDB-153 medium supplemented with 0.05 mg/mL l -histidine, 0.099 mg/mL l -isoleucine, 0.014 mg/mL l -methionine, 0.010 mg/mL l -tryptophan, 0.014 mg/mL l -tyrosine, 0.015 mg/mL l -phenylalanine, 5 μg/mL bovine insulin, and 50 ng/mL prostaglandin E1 all purchased from Sigma-Aldrich.

    Cytotoxicity Assay:

    Article Title: Engineering of near infrared fluorescent proteinoid-poly(L-lactic acid) particles for in vivo colon cancer detection
    Article Snippet: .. Materials The following analytical-grade chemicals were purchased from commercial sources and were used without further purification: L-glutamic acid (E), L-phenylalanine (F), indiocyaninegreen (ICG), human serum albumin (HSA), bovine plasma fibrinogen, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), Matrigel, Triton-x-100 and monoclonal anti-CEA antibodies (T86-66) from Sigma (Rehovot, Israel); Poly(L-lactic acid) MW 2,000 Da from Polysciences (Warrington, PA, USA); N -hydroxysulfosuccinimide (Sulfo-NHS) and 2-morpholino ethanesulfonic acid (MES, pH 6) from Thermo Fisher Scientific (Rockford, IL, USA); Phosphate Buffered Saline (PBS), Minimum Essential Medium (MEM) eagle, McCoy’s 5A medium and Dulbecco’s modification of eagle’s medium (DMEM), Fetal Bovine Serum (FBS), glutamine, penicillin, streptomycin and mycoplasma detection kit from Biological Industries (Bet Haemek, Israel); cell cytotoxicity assay kit (LDH) from Roche (Switzerland); LS174t, SW480 and HT29 cell lines from American Type Culture Collection (ATCC); donkey anti-rabbit IgG from Jackson ImmunoResearch Laboratories (West Grove, PA, USA); water was purified by passing deionized water through an Elgastat Spectrum reverse osmosis system (Elga Ltd., High Wycombe, UK). ..

    Transformation Assay:

    Article Title: Enhanced Cytotoxicity from Deoxyguanosine-Enriched T-oligo in Prostate Cancer Cells
    Article Snippet: The pZ-HPV-7 prostate epithelium cell line, transformed by the human papillomavirus 18 (HPV-18), was obtained from Dr. Tai C. Chen, Boston University School of Medicine. .. The LNCaP cell line was maintained in 10% fetal bovine serum (Hyclone); RPMI1640 medium (Gibco); 2 mM l -glutamine (Gibco); 200 U penicillin/mL; 200 μg streptomycin/mL (Gibco). pZ-HPV-7 cells were maintained in MCDB-153 medium supplemented with 0.05 mg/mL l -histidine, 0.099 mg/mL l -isoleucine, 0.014 mg/mL l -methionine, 0.010 mg/mL l -tryptophan, 0.014 mg/mL l -tyrosine, 0.015 mg/mL l -phenylalanine, 5 μg/mL bovine insulin, and 50 ng/mL prostaglandin E1 all purchased from Sigma-Aldrich.

    Binding Assay:

    Article Title: Biochemical Evaluation of Phenylalanine Ammonia Lyase from Endemic Plant Cyathobasis fruticulosa (Bunge) Aellen. for the Dietary Treatment of Phenylketonuria
    Article Snippet: Protein concentrations were determined by a dye- -binding Bradford method using Bradford reagent (Sigma- -Aldrich) and bovine serum albumin (Amresco, Cleveland, OH, USA) as the protein standard ( ). .. The enzyme reaction mixture contained 100 mM Tris- -HCl, 40 mM l -phenylalanine, ≥98% (Sigma-Aldrich), and an aliquot of the enzyme in a total volume of 1 mL at pH=8.8.

    Injection:

    Article Title: Glutamine and Leucine Provide Enhanced Protective Immunity Against Mucosal Infection with Herpes Simplex Virus Type 1
    Article Snippet: Reagents and antibodies L-Gln (G-3126), L-Leu (L8912), L-glutamic acid (Glu) (G8415), L-phenylalanine (Phe) (P5482), L-lysine (Lys) (L5501), L-asparagine (Asn) (A0884), and L-Arg were purchased from Sigma. .. All amino acids were dissolved in PBS (40mg/ml), and a volume of 200ul/mouse was administered twice daily via intra-peritoneal injection after the initial (Day 0) administration of HSV-1.

    Software:

    Article Title: Autoubiquitination of the 26S Proteasome on Rpn13 Regulates Breakdown of Ubiquitin Conjugates
    Article Snippet: To measure the degradation rate of cellular proteins, HEK293 cells were first labeled for 24 h with tritium-labeled phenylalanine (Phe L-[3,4,5 3 H], American Radiolabeled Chemicals, Inc. ART0614, stock 1 mCi/ml in 0.01 M HCl, final 5 μCi/ml) and then chased for 1 h with complete media containing 2 mM cold phenylalanine (Sigma, P5482-25G) to allow turnover of short-lived proteins. .. 200 μl supernatant after TCA precipitation was mixed with 3 ml Ultima Gold Scintillation Fluid (PerkinElmer, 6013327) and TCA-soluble radioactivity was measured with a PerkinElmer Tri-Carb 2910TR Liquid Scintillation Analyzer equipped with QuantaSmart™ software.

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  • 90
    Millipore chemotactic peptide formyl met leu phe oh fmlp
    Reduction of basal NO by AP-CAV induced significant leukocyte adhesion without increasing Lp in the absence of a secondary stimulation. A : paired video images from two rat venules. The two images on the left show the same vessel under control conditions and after AP-CAV (10 μM)-induced leukocyte adhesion. The two images on the right show that the application of sodium nitroprusside (SNP) abolished AP-CAV-induced leukocyte adhesion. B : the time course of Lp changes from a representative experiment showing that AP-CAV (10 μM)-induced leukocyte adhesion (25/100 μm of vessel length) did not increase Lp, unless <t>formyl-Met-Leu-Phe-OH</t> <t>(fMLP)</t> (10 μM) was added to the perfusate. C : summary graph showing the correlation between the number of adherent leukocytes (per 100 μm vessel length) and the changes in Lp in four group of studies. Perfusion vessels with AP-CAV at 1 μM ( n = 3) and 10 μM ( n = 5) show AP-CAV dose-dependent increases in leukocyte adhesion and fMLP-induced increases in Lp. The application of SNP attenuated AP-CAV-induced leukocyte adhesion ( n = 4), and replacing AP-CAV with scrambled AP-CAV (AP-CAV-X) showed no effect on leukocyte adhesion ( n = 3). The blank bars represent the control values. The arrows indicate the procedures of 30 min of AP-CAV or AP-CAV-X perfusion followed by 10 min of resumed blood flow. The dashed line bars represent values measured after resumed blood flow in AP-CAV or AP-CAV-X perfused vessels. *Significant increases from the control values ( P
    Chemotactic Peptide Formyl Met Leu Phe Oh Fmlp, 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/chemotactic peptide formyl met leu phe oh fmlp/product/Millipore
    Average 90 stars, based on 2 article reviews
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    81
    Millipore anti γh2ax
    Defective meiotic recombination and crossover formation in infertile Setx −/− males. A. Initiation and repair of programmed DNA DSB as shown by <t>γH2AX</t> staining of spermatocytes spreads of Setx +/+ and Setx −/− adult mice. At pachytene, γH2AX staining is restricted to the XY chromosomes (circle) in Setx +/+ spermatocytes, whereas some γH2AX foci remained on asynapsed autosomes indicating persistence of unrepaired DSB in Setx −/− . Normal γH2AX staining of the XY chromosomes (circle) was observed in both Setx +/+ and Setx −/− pachytene stage spermatocytes. Scale bar, 20 µm. XY, sex chromosomes. B. Persistence of Rad51 foci at pachytene stage in Setx −/− spermatocytes indicating the presence of unrepaired DSBs (compare 1 and 2). Scale bar, 20 µm. C. Quantitation of Rad51 foci revealed a 6-fold increase in the number of Rad51 foci at pachytene stage in Setx −/− as compared to Setx +/+ (Student's t-test, n = 50), * indicates p
    Anti γh2ax, supplied by Millipore, used in various techniques. Bioz Stars score: 81/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti γh2ax/product/Millipore
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    Image Search Results


    Reduction of basal NO by AP-CAV induced significant leukocyte adhesion without increasing Lp in the absence of a secondary stimulation. A : paired video images from two rat venules. The two images on the left show the same vessel under control conditions and after AP-CAV (10 μM)-induced leukocyte adhesion. The two images on the right show that the application of sodium nitroprusside (SNP) abolished AP-CAV-induced leukocyte adhesion. B : the time course of Lp changes from a representative experiment showing that AP-CAV (10 μM)-induced leukocyte adhesion (25/100 μm of vessel length) did not increase Lp, unless formyl-Met-Leu-Phe-OH (fMLP) (10 μM) was added to the perfusate. C : summary graph showing the correlation between the number of adherent leukocytes (per 100 μm vessel length) and the changes in Lp in four group of studies. Perfusion vessels with AP-CAV at 1 μM ( n = 3) and 10 μM ( n = 5) show AP-CAV dose-dependent increases in leukocyte adhesion and fMLP-induced increases in Lp. The application of SNP attenuated AP-CAV-induced leukocyte adhesion ( n = 4), and replacing AP-CAV with scrambled AP-CAV (AP-CAV-X) showed no effect on leukocyte adhesion ( n = 3). The blank bars represent the control values. The arrows indicate the procedures of 30 min of AP-CAV or AP-CAV-X perfusion followed by 10 min of resumed blood flow. The dashed line bars represent values measured after resumed blood flow in AP-CAV or AP-CAV-X perfused vessels. *Significant increases from the control values ( P

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: Caveolin-1 scaffolding domain promotes leukocyte adhesion by reduced basal endothelial nitric oxide-mediated ICAM-1 phosphorylation in rat mesenteric venules

    doi: 10.1152/ajpheart.00382.2013

    Figure Lengend Snippet: Reduction of basal NO by AP-CAV induced significant leukocyte adhesion without increasing Lp in the absence of a secondary stimulation. A : paired video images from two rat venules. The two images on the left show the same vessel under control conditions and after AP-CAV (10 μM)-induced leukocyte adhesion. The two images on the right show that the application of sodium nitroprusside (SNP) abolished AP-CAV-induced leukocyte adhesion. B : the time course of Lp changes from a representative experiment showing that AP-CAV (10 μM)-induced leukocyte adhesion (25/100 μm of vessel length) did not increase Lp, unless formyl-Met-Leu-Phe-OH (fMLP) (10 μM) was added to the perfusate. C : summary graph showing the correlation between the number of adherent leukocytes (per 100 μm vessel length) and the changes in Lp in four group of studies. Perfusion vessels with AP-CAV at 1 μM ( n = 3) and 10 μM ( n = 5) show AP-CAV dose-dependent increases in leukocyte adhesion and fMLP-induced increases in Lp. The application of SNP attenuated AP-CAV-induced leukocyte adhesion ( n = 4), and replacing AP-CAV with scrambled AP-CAV (AP-CAV-X) showed no effect on leukocyte adhesion ( n = 3). The blank bars represent the control values. The arrows indicate the procedures of 30 min of AP-CAV or AP-CAV-X perfusion followed by 10 min of resumed blood flow. The dashed line bars represent values measured after resumed blood flow in AP-CAV or AP-CAV-X perfused vessels. *Significant increases from the control values ( P

    Article Snippet: The chemotactic peptide formyl-Met-Leu-Phe-OH (fMLP) was purchased from Calbiochem (San Diego, CA).

    Techniques: Flow Cytometry

    Defective meiotic recombination and crossover formation in infertile Setx −/− males. A. Initiation and repair of programmed DNA DSB as shown by γH2AX staining of spermatocytes spreads of Setx +/+ and Setx −/− adult mice. At pachytene, γH2AX staining is restricted to the XY chromosomes (circle) in Setx +/+ spermatocytes, whereas some γH2AX foci remained on asynapsed autosomes indicating persistence of unrepaired DSB in Setx −/− . Normal γH2AX staining of the XY chromosomes (circle) was observed in both Setx +/+ and Setx −/− pachytene stage spermatocytes. Scale bar, 20 µm. XY, sex chromosomes. B. Persistence of Rad51 foci at pachytene stage in Setx −/− spermatocytes indicating the presence of unrepaired DSBs (compare 1 and 2). Scale bar, 20 µm. C. Quantitation of Rad51 foci revealed a 6-fold increase in the number of Rad51 foci at pachytene stage in Setx −/− as compared to Setx +/+ (Student's t-test, n = 50), * indicates p

    Journal: PLoS Genetics

    Article Title: Senataxin Plays an Essential Role with DNA Damage Response Proteins in Meiotic Recombination and Gene Silencing

    doi: 10.1371/journal.pgen.1003435

    Figure Lengend Snippet: Defective meiotic recombination and crossover formation in infertile Setx −/− males. A. Initiation and repair of programmed DNA DSB as shown by γH2AX staining of spermatocytes spreads of Setx +/+ and Setx −/− adult mice. At pachytene, γH2AX staining is restricted to the XY chromosomes (circle) in Setx +/+ spermatocytes, whereas some γH2AX foci remained on asynapsed autosomes indicating persistence of unrepaired DSB in Setx −/− . Normal γH2AX staining of the XY chromosomes (circle) was observed in both Setx +/+ and Setx −/− pachytene stage spermatocytes. Scale bar, 20 µm. XY, sex chromosomes. B. Persistence of Rad51 foci at pachytene stage in Setx −/− spermatocytes indicating the presence of unrepaired DSBs (compare 1 and 2). Scale bar, 20 µm. C. Quantitation of Rad51 foci revealed a 6-fold increase in the number of Rad51 foci at pachytene stage in Setx −/− as compared to Setx +/+ (Student's t-test, n = 50), * indicates p

    Article Snippet: Slides were incubated with anti-R-loop (1∶100, S9.6) or anti-γH2AX (1∶100, Y-P1016, Millipore) antibody overnight at 4°C in a humidified chamber.

    Techniques: Staining, Mouse Assay, Quantitation Assay

    Defective localisation and diffusion of DNA damage response proteins in Setx −/− . A. Absence of ATR diffusion over the XY chromatin domain in Setx −/− compared to Setx +/+ . Scale bar, 5 µm. B. Incomplete diffusion of MDC1 over the XY chromatin domain in Setx −/− , as indicated by the white arrow. Scale bar, 5 µm. C. Reduced intensity and diffusion of γH2AX staining on the XY chromosomes in Setx −/− compared to Setx +/+ . D. Altered XY chromosomes structure and formation in Setx −/− as shown by SCP3 staining. Scale bar 5 µm. E. Percentage of Setx +/+ and Setx −/− pachytene spermatocytes at days 16, 20 and 22 with clearly distinguishable XY chromosomes. At every time point, a significant higher percentage of distinguishable XY chromosomes was observed in Setx +/+ (p

    Journal: PLoS Genetics

    Article Title: Senataxin Plays an Essential Role with DNA Damage Response Proteins in Meiotic Recombination and Gene Silencing

    doi: 10.1371/journal.pgen.1003435

    Figure Lengend Snippet: Defective localisation and diffusion of DNA damage response proteins in Setx −/− . A. Absence of ATR diffusion over the XY chromatin domain in Setx −/− compared to Setx +/+ . Scale bar, 5 µm. B. Incomplete diffusion of MDC1 over the XY chromatin domain in Setx −/− , as indicated by the white arrow. Scale bar, 5 µm. C. Reduced intensity and diffusion of γH2AX staining on the XY chromosomes in Setx −/− compared to Setx +/+ . D. Altered XY chromosomes structure and formation in Setx −/− as shown by SCP3 staining. Scale bar 5 µm. E. Percentage of Setx +/+ and Setx −/− pachytene spermatocytes at days 16, 20 and 22 with clearly distinguishable XY chromosomes. At every time point, a significant higher percentage of distinguishable XY chromosomes was observed in Setx +/+ (p

    Article Snippet: Slides were incubated with anti-R-loop (1∶100, S9.6) or anti-γH2AX (1∶100, Y-P1016, Millipore) antibody overnight at 4°C in a humidified chamber.

    Techniques: Diffusion-based Assay, Staining

    Evidence of DNA/RNA hybrids (R-loops) accumulation in spermatocytes of ARCA mouse model. A. Histological cross-sections of seminiferous tubules from ARCA mouse models Setx −/− , Atm −/− , Aptx −/− , Tdp1 −/− and wildtype (WT) littermates. Sections were H E stained. Scale bar, 100 µm. B. Immunostaining of testes sections with R-loop (S9.6) antibody (red) and TUNEL (green). Nuclei were stained using Hoechst 33342 (blue). Scale bar, 100 µm. Magnified views of the tubules are also shown. Representative spermatocytes staining positive for both R-loop and TUNEL are indicated by white arrows. C. Immunostaining of testes sections with γH2AX (red), a marker of DNA DSBs, and TUNEL (green). Scale bar, 100 µm.

    Journal: PLoS ONE

    Article Title: R-Loops in Proliferating Cells but Not in the Brain: Implications for AOA2 and Other Autosomal Recessive Ataxias

    doi: 10.1371/journal.pone.0090219

    Figure Lengend Snippet: Evidence of DNA/RNA hybrids (R-loops) accumulation in spermatocytes of ARCA mouse model. A. Histological cross-sections of seminiferous tubules from ARCA mouse models Setx −/− , Atm −/− , Aptx −/− , Tdp1 −/− and wildtype (WT) littermates. Sections were H E stained. Scale bar, 100 µm. B. Immunostaining of testes sections with R-loop (S9.6) antibody (red) and TUNEL (green). Nuclei were stained using Hoechst 33342 (blue). Scale bar, 100 µm. Magnified views of the tubules are also shown. Representative spermatocytes staining positive for both R-loop and TUNEL are indicated by white arrows. C. Immunostaining of testes sections with γH2AX (red), a marker of DNA DSBs, and TUNEL (green). Scale bar, 100 µm.

    Article Snippet: Slides were incubated with anti-R-loop (1∶100, S9.6), anti-γH2AX (1∶100, Y-P1016, Millipore) or anti-Ki67 (1∶100, ab15580, Abcam) antibodies overnight at 4°C in a humidified chamber.

    Techniques: Staining, Immunostaining, TUNEL Assay, Marker