fmoc ala oh  (Millipore)


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    Name:
    Fmoc Ala OH
    Description:

    Catalog Number:
    531480
    Price:
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    Structured Review

    Millipore fmoc ala oh
    Fmoc Ala OH

    https://www.bioz.com/result/fmoc ala oh/product/Millipore
    Average 93 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    fmoc ala oh - by Bioz Stars, 2020-09
    93/100 stars

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    Article Title: Thiol-Selective Fluorogenic Probes for Labeling and Release
    Article Snippet: The following Fmoc-protected amino acids with side chain protecting groups were used as received from NovaBiochem: Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gly-OH, Fmoc-Ile-OH, Fmoc-Lys(Boc)-OH, Fmoc-Phe-OH, Fmoc-Thr(tBu)-OH.

    Article Title: Nanoscale physicochemical properties of chain- and step-growth polymerized PEG hydrogels affect cell-material interactions
    Article Snippet: Most amino acids including Fmoc-Ala-OH, Fmoc-Lys(Aloc)-OH, Fmoc-Val-OH, Fmoc-Ile-OH were purchased from Novabiochem; Fmoc-(PEG)6 -CH2 CH2 COOH, Fmoc-Gly-Wang Resin (100–200 mesh), Fmoc-Arg(PbF)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Gly-OH, Fmoc-Cys(Trt)-OH, Fmoc-Ser(tBu)-OH, and Fmoc-Lys(Boc)-OH were purchased from Aapptec.

    Article Title: HIV-1 Integrase-Targeted Short Peptides Derived from a Viral Protein R Sequence
    Article Snippet: Amino acid reagents, Fmoc-Gly-OH, Fmoc-Phe-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH and Fmoc-Ala-OH were obtained from NOVA Biochem (MilliporeSigma, Burlington, MA, USA).

    Article Title: Synthesis and Chromatography-Free Purification of PNA-PEO Conjugates for the Functionalisation of Gold Sensors
    Article Snippet: Fmoc-Ala-OH was from Novabiochem Merck (Darmstadt, Germany), whereas Fmoc protected PNA monomers were purchased from Polyorg (Leominster, MA, USA); α-amino-ω-carboxy-poly(ethylene glycol) (H2 N-PEO-COOH, 2 KDa and 5 KDa) was purchased from Laysan Bio (Huntsville, AL, USA), or IRIS Biotech Gmbh (Marktredwitz, Germany), 2-(1H -7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium hexafluorophosphate (HATU) and N -hydroxybenzotriazole (HOBt) were purchased from Advanced Biotech Italia (Milan, Italy).

    Article Title: Site-specific N-terminal labeling of proteins using sortase-mediated reactions
    Article Snippet: 5(6)-carboxy-tetramethylrhodamine (5(6)-TAMRA; Novabiochem, cat. no, 815030) Biotin (Sigma Aldrich, cat. no. B4501) Fmoc-Ala-OH (Novabiochem, cat. no. 852003) Fmoc-Lysine(Mtt)-OH (EMD biosciences, cat. no. 04-12-1137) Fmoc-Cys(Trt)-OH (Novabiochem, cat. no. 852008) Fmoc-Gly-OH (Novabiochem, cat. no. 852001) Fmoc-Thr(tBu)-OH (Novabiochem, cat. no. 852000) Fmoc-Glu(OtBu)-OH(Novabiochem, cat. no. 852009) Fmoc-Pro-OH (Novabiochem, cat. no. 852017) Fmo-Leu-OH (Novabiochem, cat. no. 852011) Fmoc-ε-caproic acid (Novabiochem, cat. no. 852053) 2-(1H-Benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU; Novabiochem, cat. no. 851006) !

    Droplet Countercurrent Chromatography:

    Article Title: Covalent Epitope Decoration of Carbon Electrodes using Solid Phase Peptide Synthesis
    Article Snippet: .. Chlorobenzene (99.8%, product 284513), lithium aluminum hydride (1.0 M in diethyl ether, product 212792), tetrahydrofuran (THF) (≥99.9%, product 401757), 4-(dimethylamino)pyridine (DMAP) (≥99%, product 522805), N,N′-dicyclohexylcarbodiimide (DCC) (99%, product D80002), dichloromethane (≥99.8%, product 270997), N,N′-diisopropylcarbodiimide (DIC) (99%, product D125407), 1-hydroxybenzotriazole (HOBt) hydrate (97%, product 711489), piperazine (99%, product P45907), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (98%, product 139009), trifluoroacetic acid (TFA) (99%, product T6508), piperidine (99%, product 104094), triethylamine (≥99.5%, product 471283), and Fmoc chloride (97%, product 160512), as well as amino acid monomers Fmoc-Pro-OH (monomer P, ≥99.0%, product 47636), Fmoc-Tyr(tBu)-OH (monomer T, ≥98.0%, product 47623), Fmoc-Ala-OH (monomer A, 95%, product 531480), Fmoc-Gly-OH (monomer G, ≥98.0%, product 47627), Fmoc-Val-OH (monomer V, ≥98.0%, product 47638), and Fmoc-His(Trt)-OH (monomer H, ≥98.0%, product 47639), and bovine serum albumin (BSA) (≥96%, product A7888) and phosphate buffered saline (PBS) (powder, product P3813) were obtained from Sigma-Aldrich. .. D-(+)-biotin (product 2031) was obtained from Calbiochem.

    Article Title: Isotope Dilution Mass Spectrometry for the Quantification of Sulfane Sulfurs
    Article Snippet: .. L-Alanine, thionyl chloride and Fmoc-Ala-OH-13 C3 were purchased from Sigma–Aldrich; 4-dimethylaminopyridine and HOBt were purchased from Acros Organics; DCC, methyl iodide, and piperidine were purchased from Alfa Aesar. ..

    Synthesized:

    Article Title: Small Molecule Inhibitors of the Neuropilin-1 Vascular Endothelial Growth Factor A (VEGF-A) Interaction †
    Article Snippet: .. Linear Peptide Synthesis Linear peptides were synthesized by Fmoc solid-phase synthesis using Wang or 2-chlorotrityl linkers in accordance with our previously reported methods. ( ) The resins and the amino acid derivatives, Fmoc-Abu-OH, Fmoc(Me)-Ala-OH, Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Phe-OH, and Fmoc-Pro-OH were purchased from Calbiochem Novabiochem (Nottingham, U.K.) and Bachem AG (Bubendorf, Switzerland). .. All solvents used were of HPLC-grade quality.

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    Millipore fmoc β ala oh
    Synthesis of PEGtide dendrons: (A) G1.0; and (B) G2.0-5.0. The dendrons were synthesized using <t>Fmoc</t> SPPS using following components: Fmoc-Lys(5-FAM)-OH; Fmoc-Lys(Fmoc)-OH; <t>Fmoc-β-Ala-OH;</t> and Fmoc-dPEG 6 -OH.
    Fmoc β Ala Oh, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fmoc β ala oh/product/Millipore
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fmoc β ala oh - by Bioz Stars, 2020-09
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    99
    Millipore pp2a phosphatase assay
    <t>PP2A</t> is important for B cell activation and Ig production in vitro . ( A ) Western blot analysis of PP2Aa and PP2Ac subunit expression in isolated B cells from the indicated mice. Quantification of PP2Ac expression in isolated B cells from the indicated mice. ( B ) ELISA analysis of the indicated serum Ig levels from the indicated mice (12–24 weeks old) ( n = 3 mice per group for 2 independent experiments). ( C and D ) Splenic B cells were isolated from the indicated mice and were stimulated with either CpG or anti-CD40 in the presence of IL-4 for 72 hours. ( C ) Left: ELISA analysis of the indicated Ig produced by in vitro cultured B cells with CpG and IL-4 stimulation. Middle: Dot plots represent the percentages of IgG + CD138 + plasma cells in total cultured B cells in the presence of CpG plus IL-4. Right: qPCR analysis on the expression of the indicated gene expression by the indicated B cells ( n = 3 per group for 2 independent experiments). ( D ) Left: ELISA analysis of the indicated Ig produced by in vitro cultured B cells after anti-CD40 and IL-4 stimulation. Middle: Dot plots indicate the percentages of IgG + CD138 + plasma cells in total B cells cultured with anti-CD40 and IL-4. Right: qPCR analysis of indicated genes in the indicated B cells ( n = 3 per group for 2 independent experiments). Paired t test, mean ± SEM.
    Pp2a Phosphatase Assay, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pp2a phosphatase assay/product/Millipore
    Average 99 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    pp2a phosphatase assay - by Bioz Stars, 2020-09
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    Synthesis of PEGtide dendrons: (A) G1.0; and (B) G2.0-5.0. The dendrons were synthesized using Fmoc SPPS using following components: Fmoc-Lys(5-FAM)-OH; Fmoc-Lys(Fmoc)-OH; Fmoc-β-Ala-OH; and Fmoc-dPEG 6 -OH.

    Journal: Bioconjugate chemistry

    Article Title: Novel monodisperse PEGtide dendrons: design, fabrication and evaluation of mannose receptor-mediated macrophage targeting

    doi: 10.1021/bc400011v

    Figure Lengend Snippet: Synthesis of PEGtide dendrons: (A) G1.0; and (B) G2.0-5.0. The dendrons were synthesized using Fmoc SPPS using following components: Fmoc-Lys(5-FAM)-OH; Fmoc-Lys(Fmoc)-OH; Fmoc-β-Ala-OH; and Fmoc-dPEG 6 -OH.

    Article Snippet: Fmoc- N -amido-dPEG® 6 -acid (MW 575.65 Da, Fmoc-dPEG6 -OH) was purchased from Quanta Biodesign Ltd. (Powell, OH); Fmoc-β-Ala-Wang Resin, Fmoc-Lys(Fmoc)-OH, Fmoc-β-Ala-OH, benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBOP) were purchased from EMD Chemicals (Gibbstown, NJ); N -α-Fmoc- N -ε-(5-carboxyfluorescein)-L-lysine (Fmoc-Lys(5-FAM)-OH) and 4”,6-diamidino-2-phenylindole, dihydrochloride (DAPI) were purchased from Anaspec (Fremont, CA); 1-hydroxybenzotrizole (HOBt) was purchased from Chem-Impex International (Wood Dale, IL); piperidine, α-D-mannopyranosylphenyl isothiocyanate, triisopropylsilane, sinapinic acid and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO); N,N -dimethylformamide (DMF), acetic anhydride, N,N -diisopropylethylamine (DIPEA) were purchased from Acros Organics (Morris Plains, NJ); trifluoroacetic acid (TFA) was purchased from Fisher Scientific (Pittsburgh, PA); diethyl ether was purchased from Honeywell Brudick & Jackson (Muskegon, MI); and PD-10 column was purchased from GE Healthcare (Piscataway, NJ).

    Techniques: Synthesized

    PP2A is important for B cell activation and Ig production in vitro . ( A ) Western blot analysis of PP2Aa and PP2Ac subunit expression in isolated B cells from the indicated mice. Quantification of PP2Ac expression in isolated B cells from the indicated mice. ( B ) ELISA analysis of the indicated serum Ig levels from the indicated mice (12–24 weeks old) ( n = 3 mice per group for 2 independent experiments). ( C and D ) Splenic B cells were isolated from the indicated mice and were stimulated with either CpG or anti-CD40 in the presence of IL-4 for 72 hours. ( C ) Left: ELISA analysis of the indicated Ig produced by in vitro cultured B cells with CpG and IL-4 stimulation. Middle: Dot plots represent the percentages of IgG + CD138 + plasma cells in total cultured B cells in the presence of CpG plus IL-4. Right: qPCR analysis on the expression of the indicated gene expression by the indicated B cells ( n = 3 per group for 2 independent experiments). ( D ) Left: ELISA analysis of the indicated Ig produced by in vitro cultured B cells after anti-CD40 and IL-4 stimulation. Middle: Dot plots indicate the percentages of IgG + CD138 + plasma cells in total B cells cultured with anti-CD40 and IL-4. Right: qPCR analysis of indicated genes in the indicated B cells ( n = 3 per group for 2 independent experiments). Paired t test, mean ± SEM.

    Journal: JCI Insight

    Article Title: Serine/threonine phosphatase PP2A is essential for optimal B cell function

    doi: 10.1172/jci.insight.130655

    Figure Lengend Snippet: PP2A is important for B cell activation and Ig production in vitro . ( A ) Western blot analysis of PP2Aa and PP2Ac subunit expression in isolated B cells from the indicated mice. Quantification of PP2Ac expression in isolated B cells from the indicated mice. ( B ) ELISA analysis of the indicated serum Ig levels from the indicated mice (12–24 weeks old) ( n = 3 mice per group for 2 independent experiments). ( C and D ) Splenic B cells were isolated from the indicated mice and were stimulated with either CpG or anti-CD40 in the presence of IL-4 for 72 hours. ( C ) Left: ELISA analysis of the indicated Ig produced by in vitro cultured B cells with CpG and IL-4 stimulation. Middle: Dot plots represent the percentages of IgG + CD138 + plasma cells in total cultured B cells in the presence of CpG plus IL-4. Right: qPCR analysis on the expression of the indicated gene expression by the indicated B cells ( n = 3 per group for 2 independent experiments). ( D ) Left: ELISA analysis of the indicated Ig produced by in vitro cultured B cells after anti-CD40 and IL-4 stimulation. Middle: Dot plots indicate the percentages of IgG + CD138 + plasma cells in total B cells cultured with anti-CD40 and IL-4. Right: qPCR analysis of indicated genes in the indicated B cells ( n = 3 per group for 2 independent experiments). Paired t test, mean ± SEM.

    Article Snippet: PP2A phosphatase assay.B cells were isolated from mouse spleens, and PP2A phosphatase assay was performed as manufactory’s instructions (MilliporeSigma, catalog 17-313).

    Techniques: Activation Assay, In Vitro, Western Blot, Expressing, Isolation, Mouse Assay, Enzyme-linked Immunosorbent Assay, Produced, Cell Culture, Real-time Polymerase Chain Reaction

    PP2A in B cells inhibits mitochondrial respiration by suppressing the expression of purine nucleoside phosphorylase (PNP). ( A ) Dot plots show mitochondrial respiration in Ppp2r1a- deficient mouse splenic B cells compared with WT mouse splenic B cells (maximal oxygen consumption rate [OCR]and space capacity OCR) ( n = 4 independent experiments). ( B ) Joint analysis of RNA sequencing and metabolomics in B cells from flox/flox mice compared with control mice ( n = 3 mice per group). ( C ) Increased PNP mRNA expression in B cells from flox/flox mice compared with control mice ( n = 6 mice per group for qPCR experiment). ( D ) Western blot analysis on PNP expression in mouse splenic B cells with PP2Aa deficiency compared with WT controls. Quantification of Western blots. ( E ) Human primary B cells were enriched from peripheral blood mononuclear cells (PBMC). Left: Representation of oxygen consumption rate (OCR) of human primary B cells exposed to the indicated treatments (9-Deazaguanine was applied as PNP inhibitor). Right: Dot plots indicate mitochondrial respiration in human primary B cells with PP2Aa deficiency compared with controls exposed to the indicated treatments. Paired t test, mean ± SEM.

    Journal: JCI Insight

    Article Title: Serine/threonine phosphatase PP2A is essential for optimal B cell function

    doi: 10.1172/jci.insight.130655

    Figure Lengend Snippet: PP2A in B cells inhibits mitochondrial respiration by suppressing the expression of purine nucleoside phosphorylase (PNP). ( A ) Dot plots show mitochondrial respiration in Ppp2r1a- deficient mouse splenic B cells compared with WT mouse splenic B cells (maximal oxygen consumption rate [OCR]and space capacity OCR) ( n = 4 independent experiments). ( B ) Joint analysis of RNA sequencing and metabolomics in B cells from flox/flox mice compared with control mice ( n = 3 mice per group). ( C ) Increased PNP mRNA expression in B cells from flox/flox mice compared with control mice ( n = 6 mice per group for qPCR experiment). ( D ) Western blot analysis on PNP expression in mouse splenic B cells with PP2Aa deficiency compared with WT controls. Quantification of Western blots. ( E ) Human primary B cells were enriched from peripheral blood mononuclear cells (PBMC). Left: Representation of oxygen consumption rate (OCR) of human primary B cells exposed to the indicated treatments (9-Deazaguanine was applied as PNP inhibitor). Right: Dot plots indicate mitochondrial respiration in human primary B cells with PP2Aa deficiency compared with controls exposed to the indicated treatments. Paired t test, mean ± SEM.

    Article Snippet: PP2A phosphatase assay.B cells were isolated from mouse spleens, and PP2A phosphatase assay was performed as manufactory’s instructions (MilliporeSigma, catalog 17-313).

    Techniques: Expressing, RNA Sequencing Assay, Mouse Assay, Real-time Polymerase Chain Reaction, Western Blot

    Increased PP2A expression and enhanced function in activated B cells or B cells from lupus-prone mice and SLE patients. ( A ) Western blot analysis on the expression of both scaffold (PP2A A ) and catalytic (PP2A C ) subunits in splenic B cells isolated from the indicated mice. Quantification of Western blots ( n = 3 per experiment, a representative experiment and pooled densitometry of the 3 experiments are shown). ( B ) Germinal center B cells (GC, CD19 + FAS + GL7 + ), plasma cells (PC, CD19 + IgG + CD138 hi ), marginal zone B cells (MZ, CD19 + CD21 + CD23 lo ), and follicular B cells (FO, CD19 + CD21 lo CD23 hi ) were FACS sorted for qPCR. Dot plots show the expression of PP2A A , and elevated expression was observed in GC and PC compared with MZ and FO. ( C ) Flow cytometry analysis of the expression of PP2Ac (indicated by mean florescence intensity, MFI) in total B cells from patients with SLE and matched healthy controls. ( D ) Dot plots show the increased MFI of PP2Ac in DN B cells (CD19 + IgD – CD27 – ), memory B cells (CD19 + IgD – CD27 + ), and plasma B cells (CD19 + IgG + CD138 + ) compared with naive B cells (CD19 + IgD + CD27 – ). ( E–H ) CD19 + human B cells were enriched by magnetic beads and cultured with the indicated stimuli for the indicated time. ( E ) Dot plots show the expression of PP2Aa in B cells stimulated with either CpG or anti-CD40 compared with unstimulated B cells 3 hours after stimulation. ( F ) Dot plots show the expression of PP2Aa in B cells stimulated with either CpG or anti-CD40 compared with unstimulated B cells 3 hours after stimulation. ( G ) Western blot analysis of the expression of PP2Aa and PP2Ac in human B cells stimulated with either CpG or anti-CD40 for 48 hours. ( H ) Dot plots show the quantification of PP2Aa and expression PP2Ac in human B cells stimulated with either CpG or anti-CD40 compared with control unstimulated B cells 3 hours after stimulation. ( I and J ) PP2A phosphatase activity was quantified using a kit from R D. The activity of PP2A is presented as the rate of phosphate release (pmol × 10 2 ). ( I ) Bar graph shows the PP2A phosphatase activity in ex vivo splenic B cells from lupus-prone Mrl.lpr mice compared with matching control Mrl.mpj mice (data pooled from 3 independent experiments, paired t test, mean ± SEM). ( J ) Splenic B cells were enriched from C57BL/6J WT mice by MACS and stimulated with the indicated reagents for several time periods; kinetic curves show the PP2A phosphatase activity ( n = 3 for 3 independent experiments, 2-way ANOVA analysis, mean ± SEM).

    Journal: JCI Insight

    Article Title: Serine/threonine phosphatase PP2A is essential for optimal B cell function

    doi: 10.1172/jci.insight.130655

    Figure Lengend Snippet: Increased PP2A expression and enhanced function in activated B cells or B cells from lupus-prone mice and SLE patients. ( A ) Western blot analysis on the expression of both scaffold (PP2A A ) and catalytic (PP2A C ) subunits in splenic B cells isolated from the indicated mice. Quantification of Western blots ( n = 3 per experiment, a representative experiment and pooled densitometry of the 3 experiments are shown). ( B ) Germinal center B cells (GC, CD19 + FAS + GL7 + ), plasma cells (PC, CD19 + IgG + CD138 hi ), marginal zone B cells (MZ, CD19 + CD21 + CD23 lo ), and follicular B cells (FO, CD19 + CD21 lo CD23 hi ) were FACS sorted for qPCR. Dot plots show the expression of PP2A A , and elevated expression was observed in GC and PC compared with MZ and FO. ( C ) Flow cytometry analysis of the expression of PP2Ac (indicated by mean florescence intensity, MFI) in total B cells from patients with SLE and matched healthy controls. ( D ) Dot plots show the increased MFI of PP2Ac in DN B cells (CD19 + IgD – CD27 – ), memory B cells (CD19 + IgD – CD27 + ), and plasma B cells (CD19 + IgG + CD138 + ) compared with naive B cells (CD19 + IgD + CD27 – ). ( E–H ) CD19 + human B cells were enriched by magnetic beads and cultured with the indicated stimuli for the indicated time. ( E ) Dot plots show the expression of PP2Aa in B cells stimulated with either CpG or anti-CD40 compared with unstimulated B cells 3 hours after stimulation. ( F ) Dot plots show the expression of PP2Aa in B cells stimulated with either CpG or anti-CD40 compared with unstimulated B cells 3 hours after stimulation. ( G ) Western blot analysis of the expression of PP2Aa and PP2Ac in human B cells stimulated with either CpG or anti-CD40 for 48 hours. ( H ) Dot plots show the quantification of PP2Aa and expression PP2Ac in human B cells stimulated with either CpG or anti-CD40 compared with control unstimulated B cells 3 hours after stimulation. ( I and J ) PP2A phosphatase activity was quantified using a kit from R D. The activity of PP2A is presented as the rate of phosphate release (pmol × 10 2 ). ( I ) Bar graph shows the PP2A phosphatase activity in ex vivo splenic B cells from lupus-prone Mrl.lpr mice compared with matching control Mrl.mpj mice (data pooled from 3 independent experiments, paired t test, mean ± SEM). ( J ) Splenic B cells were enriched from C57BL/6J WT mice by MACS and stimulated with the indicated reagents for several time periods; kinetic curves show the PP2A phosphatase activity ( n = 3 for 3 independent experiments, 2-way ANOVA analysis, mean ± SEM).

    Article Snippet: PP2A phosphatase assay.B cells were isolated from mouse spleens, and PP2A phosphatase assay was performed as manufactory’s instructions (MilliporeSigma, catalog 17-313).

    Techniques: Expressing, Mouse Assay, Western Blot, Isolation, FACS, Real-time Polymerase Chain Reaction, Flow Cytometry, Magnetic Beads, Cell Culture, Activity Assay, Ex Vivo, Magnetic Cell Separation

    PP2A is critical for B cell activation, differentiation, and Ig production in vivo. ( A ) Dot plots show the percentages of spontaneous germinal center B cells (GC-CD19 + FAS + PNA + ) and T follicular helper cells (Tfh-CD4 + CXCR5 hi PD1 hi ) in spleens of the indicated 24-week-old mice ( n = 6 mice per group for 2 independent experiments). ( B and C ) Mice were immunized i.p. with 0.2 mL/mouse SRBC. ( B ) IHC staining of frozen spleen sections of the indicated mice ( n = 3 mice per group in 2 independent experiments). Bar graphs show the quantification (percentage) of follicles with germinal centers in total splenic follicles from the indicated mice ( n = 3 mice per group in 2 independent experiments). ( C ) Flow cytometry analysis (%) of germinal center B cells (GC-CD19 + FAS + PNA + ), T follicular helper cells (Tfh-CD4 + CXCR5 hi PD1 hi ), and IgG plasma cells (PC-CD19 + IgG + CD138 + ) in the spleens of the indicated mice after SRBC immunization. ( D and E ) Indicated mice were i.p. immunized with 100 μg/mouse NP-Ficoll for 5 days or 100 μg/mouse NP-CGG in 5% alum for 14 days. ( D ) ELISA analysis for high affinity (NP-7) or low affinity (NP-41) for NP antigen-specific IgM in the serum of 12-week-old flox/flox mice compared with control mice 5 days after immunization with T-independent antigen NP-Ficoll ( n = 3 per group in 2 independent experiments). ( E ) ELISA analysis of high affinity (NP-7) or all affinity (NP-41) NP antigen-specific IgG in the serum of 12-week-old flox/flox mice compared with control mice 14 days after immunization with T-dependent antigen NP-CGG ( n = 3 per group in 2 independent experiments). Paired t test, ** P

    Journal: JCI Insight

    Article Title: Serine/threonine phosphatase PP2A is essential for optimal B cell function

    doi: 10.1172/jci.insight.130655

    Figure Lengend Snippet: PP2A is critical for B cell activation, differentiation, and Ig production in vivo. ( A ) Dot plots show the percentages of spontaneous germinal center B cells (GC-CD19 + FAS + PNA + ) and T follicular helper cells (Tfh-CD4 + CXCR5 hi PD1 hi ) in spleens of the indicated 24-week-old mice ( n = 6 mice per group for 2 independent experiments). ( B and C ) Mice were immunized i.p. with 0.2 mL/mouse SRBC. ( B ) IHC staining of frozen spleen sections of the indicated mice ( n = 3 mice per group in 2 independent experiments). Bar graphs show the quantification (percentage) of follicles with germinal centers in total splenic follicles from the indicated mice ( n = 3 mice per group in 2 independent experiments). ( C ) Flow cytometry analysis (%) of germinal center B cells (GC-CD19 + FAS + PNA + ), T follicular helper cells (Tfh-CD4 + CXCR5 hi PD1 hi ), and IgG plasma cells (PC-CD19 + IgG + CD138 + ) in the spleens of the indicated mice after SRBC immunization. ( D and E ) Indicated mice were i.p. immunized with 100 μg/mouse NP-Ficoll for 5 days or 100 μg/mouse NP-CGG in 5% alum for 14 days. ( D ) ELISA analysis for high affinity (NP-7) or low affinity (NP-41) for NP antigen-specific IgM in the serum of 12-week-old flox/flox mice compared with control mice 5 days after immunization with T-independent antigen NP-Ficoll ( n = 3 per group in 2 independent experiments). ( E ) ELISA analysis of high affinity (NP-7) or all affinity (NP-41) NP antigen-specific IgG in the serum of 12-week-old flox/flox mice compared with control mice 14 days after immunization with T-dependent antigen NP-CGG ( n = 3 per group in 2 independent experiments). Paired t test, ** P

    Article Snippet: PP2A phosphatase assay.B cells were isolated from mouse spleens, and PP2A phosphatase assay was performed as manufactory’s instructions (MilliporeSigma, catalog 17-313).

    Techniques: Activation Assay, In Vivo, Mouse Assay, Immunohistochemistry, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay

    PP2A expression in B cells for disease development in mice after injection of pristine. ( A and B ) Indicated mice were challenged i.p. with 0.5 mL pristine. ( A ) Blot graph shows ANA antibody levels in the serum of flox/flox and control mice 1 month after pristine challenge ( n = 6 mice per group). ( B ) Immunofluorescence image analysis to detect IgG deposition in the kidneys of flox/flox and control mice 6 months after pristine challenge ( n = 6 mice per group). ( C ) Western blot analysis on pSTAT3 and total STAT3 expression in B cells with PP2A deficiency. Quantification of Western blots. ( D–F ) Human B cells were enriched from peripheral blood mononuclear cells (PBMC) using MACS cell separation kits. ( D ) Western blot analysis to detect PP2Aa and PP2Ac in human B cells treated with control or PP2Aa siRNA. Quantification of Western blot densities. ( E ) Flow cytometry analysis on pSTAT3 induction during B cell activation in the absence of PP2Aa. ( F ) Immunofluorescence image analysis on pSTAT3 expression during B cell activation in the absence of PP2Aa. Original magnification ×63. Paired t test, * P

    Journal: JCI Insight

    Article Title: Serine/threonine phosphatase PP2A is essential for optimal B cell function

    doi: 10.1172/jci.insight.130655

    Figure Lengend Snippet: PP2A expression in B cells for disease development in mice after injection of pristine. ( A and B ) Indicated mice were challenged i.p. with 0.5 mL pristine. ( A ) Blot graph shows ANA antibody levels in the serum of flox/flox and control mice 1 month after pristine challenge ( n = 6 mice per group). ( B ) Immunofluorescence image analysis to detect IgG deposition in the kidneys of flox/flox and control mice 6 months after pristine challenge ( n = 6 mice per group). ( C ) Western blot analysis on pSTAT3 and total STAT3 expression in B cells with PP2A deficiency. Quantification of Western blots. ( D–F ) Human B cells were enriched from peripheral blood mononuclear cells (PBMC) using MACS cell separation kits. ( D ) Western blot analysis to detect PP2Aa and PP2Ac in human B cells treated with control or PP2Aa siRNA. Quantification of Western blot densities. ( E ) Flow cytometry analysis on pSTAT3 induction during B cell activation in the absence of PP2Aa. ( F ) Immunofluorescence image analysis on pSTAT3 expression during B cell activation in the absence of PP2Aa. Original magnification ×63. Paired t test, * P

    Article Snippet: PP2A phosphatase assay.B cells were isolated from mouse spleens, and PP2A phosphatase assay was performed as manufactory’s instructions (MilliporeSigma, catalog 17-313).

    Techniques: Expressing, Mouse Assay, Injection, Immunofluorescence, Western Blot, Magnetic Cell Separation, Flow Cytometry, Activation Assay

    PP2A counters smoke-induced inflammation in mice. (A) β-galactosidase immunostains were conducted on lung sections from mice injected intratracheally with 2 μg of β-galactosidase protein or PBS (control). (B) Intratracheal transfection

    Journal: Toxicological Sciences

    Article Title: Protein Phosphatase 2A Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung

    doi: 10.1093/toxsci/kfr351

    Figure Lengend Snippet: PP2A counters smoke-induced inflammation in mice. (A) β-galactosidase immunostains were conducted on lung sections from mice injected intratracheally with 2 μg of β-galactosidase protein or PBS (control). (B) Intratracheal transfection

    Article Snippet: PP2A activity was determined by immunoprecipitating the catalytic subunit of PP2A from the tissue or cell culture samples and then measuring the phosphatase activity of the immunoprecipitated protein using a specific PP2A assay (17-313, Millipore).

    Techniques: Mouse Assay, Injection, Transfection

    Decreasing PP2A exacerbates IL-8 and MMP-1 production in smoke-treated SAE cells. (A) Western blot for PPP2R1A was done on protein from SAE cells stably transfected with control or PPP2R1A shRNA. (B) PP2A activity assays were conducted on control- or

    Journal: Toxicological Sciences

    Article Title: Protein Phosphatase 2A Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung

    doi: 10.1093/toxsci/kfr351

    Figure Lengend Snippet: Decreasing PP2A exacerbates IL-8 and MMP-1 production in smoke-treated SAE cells. (A) Western blot for PPP2R1A was done on protein from SAE cells stably transfected with control or PPP2R1A shRNA. (B) PP2A activity assays were conducted on control- or

    Article Snippet: PP2A activity was determined by immunoprecipitating the catalytic subunit of PP2A from the tissue or cell culture samples and then measuring the phosphatase activity of the immunoprecipitated protein using a specific PP2A assay (17-313, Millipore).

    Techniques: Western Blot, Stable Transfection, Transfection, shRNA, Activity Assay

    Increasing PP2A activity deters cytokine and protease expression in SAE cells. (A) PP2A activity was measured in SAE cells transfected with PP2A protein in 20 μl of Pro-Ject transfection reagent. Activity is expressed as picomole phosphate liberated

    Journal: Toxicological Sciences

    Article Title: Protein Phosphatase 2A Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung

    doi: 10.1093/toxsci/kfr351

    Figure Lengend Snippet: Increasing PP2A activity deters cytokine and protease expression in SAE cells. (A) PP2A activity was measured in SAE cells transfected with PP2A protein in 20 μl of Pro-Ject transfection reagent. Activity is expressed as picomole phosphate liberated

    Article Snippet: PP2A activity was determined by immunoprecipitating the catalytic subunit of PP2A from the tissue or cell culture samples and then measuring the phosphatase activity of the immunoprecipitated protein using a specific PP2A assay (17-313, Millipore).

    Techniques: Activity Assay, Expressing, Transfection

    Inhibiting PP2A enhances acute smoke–induced lung inflammation. (A) Mice were exposed to cigarette smoke following daily injections of okadaic acid (2 μg/kg ip) for 3 days. Mice were euthanized on day 3 ( n = 5 for each group). (B) PP2A

    Journal: Toxicological Sciences

    Article Title: Protein Phosphatase 2A Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung

    doi: 10.1093/toxsci/kfr351

    Figure Lengend Snippet: Inhibiting PP2A enhances acute smoke–induced lung inflammation. (A) Mice were exposed to cigarette smoke following daily injections of okadaic acid (2 μg/kg ip) for 3 days. Mice were euthanized on day 3 ( n = 5 for each group). (B) PP2A

    Article Snippet: PP2A activity was determined by immunoprecipitating the catalytic subunit of PP2A from the tissue or cell culture samples and then measuring the phosphatase activity of the immunoprecipitated protein using a specific PP2A assay (17-313, Millipore).

    Techniques: Mouse Assay

    PP2A activity affects JNK activation in the lung. (A and B) Western blots for pJNK, JNK, and actin were conducted on lung protein from mice at baseline, 2 h (A) or 24 h (B) postsmoke exposure. (C) AP-1 activation assays were conducted on nuclear protein

    Journal: Toxicological Sciences

    Article Title: Protein Phosphatase 2A Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung

    doi: 10.1093/toxsci/kfr351

    Figure Lengend Snippet: PP2A activity affects JNK activation in the lung. (A and B) Western blots for pJNK, JNK, and actin were conducted on lung protein from mice at baseline, 2 h (A) or 24 h (B) postsmoke exposure. (C) AP-1 activation assays were conducted on nuclear protein

    Article Snippet: PP2A activity was determined by immunoprecipitating the catalytic subunit of PP2A from the tissue or cell culture samples and then measuring the phosphatase activity of the immunoprecipitated protein using a specific PP2A assay (17-313, Millipore).

    Techniques: Activity Assay, Activation Assay, Western Blot, Mouse Assay

    Cigarette smoke activates PP2A in human SAE cells. (A) PP2A activity assays were conducted on control (white bars) or CSE-treated (black bars) SAE cells ( n = 4). Graphs are represented as mean ± SEM. p values shown, comparing both treatments connected

    Journal: Toxicological Sciences

    Article Title: Protein Phosphatase 2A Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung

    doi: 10.1093/toxsci/kfr351

    Figure Lengend Snippet: Cigarette smoke activates PP2A in human SAE cells. (A) PP2A activity assays were conducted on control (white bars) or CSE-treated (black bars) SAE cells ( n = 4). Graphs are represented as mean ± SEM. p values shown, comparing both treatments connected

    Article Snippet: PP2A activity was determined by immunoprecipitating the catalytic subunit of PP2A from the tissue or cell culture samples and then measuring the phosphatase activity of the immunoprecipitated protein using a specific PP2A assay (17-313, Millipore).

    Techniques: Activity Assay

    Cigarette smoke activates PP2A in mouse lungs in vivo . (A) PP2A and (B) NF-κB activities were measured in the lungs of wild-type nonexposed (white bars) and various times of smoke-exposed mice (black bars) ( n = 5 for each group). (C) Quantitative

    Journal: Toxicological Sciences

    Article Title: Protein Phosphatase 2A Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung

    doi: 10.1093/toxsci/kfr351

    Figure Lengend Snippet: Cigarette smoke activates PP2A in mouse lungs in vivo . (A) PP2A and (B) NF-κB activities were measured in the lungs of wild-type nonexposed (white bars) and various times of smoke-exposed mice (black bars) ( n = 5 for each group). (C) Quantitative

    Article Snippet: PP2A activity was determined by immunoprecipitating the catalytic subunit of PP2A from the tissue or cell culture samples and then measuring the phosphatase activity of the immunoprecipitated protein using a specific PP2A assay (17-313, Millipore).

    Techniques: In Vivo, Mouse Assay

    PP2A activity is increased in human emphysema. (A) PP2A activity was measured in lung tissue from age-matched normal controls ( n = 9) and emphysema subjects ( n = 10) using a specific PP2A phosphatase activity assay. Graph is represented as mean ±

    Journal: Toxicological Sciences

    Article Title: Protein Phosphatase 2A Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung

    doi: 10.1093/toxsci/kfr351

    Figure Lengend Snippet: PP2A activity is increased in human emphysema. (A) PP2A activity was measured in lung tissue from age-matched normal controls ( n = 9) and emphysema subjects ( n = 10) using a specific PP2A phosphatase activity assay. Graph is represented as mean ±

    Article Snippet: PP2A activity was determined by immunoprecipitating the catalytic subunit of PP2A from the tissue or cell culture samples and then measuring the phosphatase activity of the immunoprecipitated protein using a specific PP2A assay (17-313, Millipore).

    Techniques: Activity Assay, Phosphatase Assay