gtp beads  (Jena Bioscience)


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  • 90
    Name:
    GTPγS
    Description:

    Catalog Number:
    NU-412-10
    Price:
    209.4
    Category:
    Nucleotides Nucleosides
    Size:
    10 mg
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    Structured Review

    Jena Bioscience gtp beads

    https://www.bioz.com/result/gtp beads/product/Jena Bioscience
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gtp beads - by Bioz Stars, 2021-04
    90/100 stars

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    Chloramphenicol Acetyltransferase Assay:

    Article Title: Phosphorylation of LRRK2 by casein kinase 1α regulates trans-Golgi clustering via differential interaction with ARHGEF7
    Article Snippet: HEK293FT cells (Invitrogen) were used for transient transfections, and for stable transfections we used with 293T stably expressing 3xFLAG-LRRK2 , (generous gift from Dr. Jean-Marc Taymans, KU, Leuven, Belgium). .. Chemicals and antibodies Chemicals and reagents used in this study were; LRRK2-IN1 (Tocris Bioscience Cat# 4273), IC261 (Millipore Cat# 400090), D4476 (Abcam Cat# ab120220), TMCB (Millipore Cat# 218718), GTP (Sigma CatG8877), ATP (Sigma Cat# A6419), protease inhibitor (Roche Cat# 04693159001), phosphatase inhibitor (Thermo Scientific Cat# 78427), GTP-beads (Jena Bioscience Cat # NU-412-10), EZview Red Protein G Affinity Gel (Sigma Cat# E3403), EZview Red ANTI-FLAG M2 Affinity Gel (Sigma Cat# F2426), 3xFlag peptide (Sigma Cat# F4799), 10x TBS (KD Medical), and 4x SDS sample buffer (Invitrogen Cat# NP0008). .. Antibodies used for western blotting (WB) or immunocytochemistry (IC) were: anti-pS935 (University of Dundee, Scotland; WB 1µg ml−1 ), anti-pS935 (Abcam Cat # ab133450; WB 1:1000), anti-pS910 (Abcam Cat # ab133449; WB 1:1000), anti-pT1410 (Abcam Cat # ab140107; WB 1:1000), anti-pT1503 (Abcam Cat # ab154423; WB 1:1000), anti-LRRK2 (Epitomics Cat# 3514-1; 1:5000), anti-LRRK2 (Millipore Cat# MABN40; WB 1:1000), anti-CK1α (Santa Cruz Cat# sc-6477; WB 1:2000), anti-CK1δ (Santa Cruz Cat# sc-55553; WB 1:1000), anti-CK1ε (Santa Cruz Cat# sc-6471; WB 1:1000), anti-CK1γ1 (Santa Cruz Cat# sc-18493; WB 1:1000) anti-ARHGEF7 (Protein Tech Cat# 14092-1-AP; WB 1:1000), anti-Hsp90 (Cell Signalling Cat# 4877; WB 1:2000), anti-alpha-tubulin (Cell Signalling Cat# 2125; WB 1:1000), anti-14-3-3 (Santa Cruz Cat# sc-629; WB 1:2000), anti-cdc37 (Cell Signalling Cat# 610576; WB 1:1000), anti-TGN46 (AbD Serotec Cat# AHP500G; IC 1:200), anti-actin (Sigma Cat# A5441; WB 1:2000), anti-FLAG (Sigma Cat# F1804; WB 1:5000, IC 1:500), anti-myc (Roche Cat# 11667149001; WB 1:2000, IC 1:500), HRP- conjugated secondary antibodies (Jackson Immunology; WB 1:5000) and fluorescent-conjugated secondary antibodies (Jackson Immunology; WB 1:50000, Life Technologies; IC 1:500).

    Protease Inhibitor:

    Article Title: Phosphorylation of LRRK2 by casein kinase 1α regulates trans-Golgi clustering via differential interaction with ARHGEF7
    Article Snippet: HEK293FT cells (Invitrogen) were used for transient transfections, and for stable transfections we used with 293T stably expressing 3xFLAG-LRRK2 , (generous gift from Dr. Jean-Marc Taymans, KU, Leuven, Belgium). .. Chemicals and antibodies Chemicals and reagents used in this study were; LRRK2-IN1 (Tocris Bioscience Cat# 4273), IC261 (Millipore Cat# 400090), D4476 (Abcam Cat# ab120220), TMCB (Millipore Cat# 218718), GTP (Sigma CatG8877), ATP (Sigma Cat# A6419), protease inhibitor (Roche Cat# 04693159001), phosphatase inhibitor (Thermo Scientific Cat# 78427), GTP-beads (Jena Bioscience Cat # NU-412-10), EZview Red Protein G Affinity Gel (Sigma Cat# E3403), EZview Red ANTI-FLAG M2 Affinity Gel (Sigma Cat# F2426), 3xFlag peptide (Sigma Cat# F4799), 10x TBS (KD Medical), and 4x SDS sample buffer (Invitrogen Cat# NP0008). .. Antibodies used for western blotting (WB) or immunocytochemistry (IC) were: anti-pS935 (University of Dundee, Scotland; WB 1µg ml−1 ), anti-pS935 (Abcam Cat # ab133450; WB 1:1000), anti-pS910 (Abcam Cat # ab133449; WB 1:1000), anti-pT1410 (Abcam Cat # ab140107; WB 1:1000), anti-pT1503 (Abcam Cat # ab154423; WB 1:1000), anti-LRRK2 (Epitomics Cat# 3514-1; 1:5000), anti-LRRK2 (Millipore Cat# MABN40; WB 1:1000), anti-CK1α (Santa Cruz Cat# sc-6477; WB 1:2000), anti-CK1δ (Santa Cruz Cat# sc-55553; WB 1:1000), anti-CK1ε (Santa Cruz Cat# sc-6471; WB 1:1000), anti-CK1γ1 (Santa Cruz Cat# sc-18493; WB 1:1000) anti-ARHGEF7 (Protein Tech Cat# 14092-1-AP; WB 1:1000), anti-Hsp90 (Cell Signalling Cat# 4877; WB 1:2000), anti-alpha-tubulin (Cell Signalling Cat# 2125; WB 1:1000), anti-14-3-3 (Santa Cruz Cat# sc-629; WB 1:2000), anti-cdc37 (Cell Signalling Cat# 610576; WB 1:1000), anti-TGN46 (AbD Serotec Cat# AHP500G; IC 1:200), anti-actin (Sigma Cat# A5441; WB 1:2000), anti-FLAG (Sigma Cat# F1804; WB 1:5000, IC 1:500), anti-myc (Roche Cat# 11667149001; WB 1:2000, IC 1:500), HRP- conjugated secondary antibodies (Jackson Immunology; WB 1:5000) and fluorescent-conjugated secondary antibodies (Jackson Immunology; WB 1:50000, Life Technologies; IC 1:500).

    Article Title: The G2385R risk factor for Parkinson’s disease enhances CHIP-dependent intracellular degradation of LRRK2
    Article Snippet: Point mutations were introduced by using the QuikChange II XL Site-Directed Mutagenesis Kit (Stratagene). .. Chemicals and reagents used in the present study were GTP (Sigma, Cat# G8877), protease inhibitor cocktail (Roche, Cat# 04693159001), HALT phosphatase inhibitor cocktail (Thermo Scientific, Cat# 78427), Geldanamycin (Sigma, Cat# G3381), GTP beads (Jena Bioscience, Cat# NU-412–10), EZview Red Protein G Affinity Gel (Sigma, Cat# E3403), EZview Red ANTI-FLAG M2 Affinity Gel (Sigma, Cat# F2426), HaloTag® Biotin Ligand (Promega, Cat# G8281), ON-TARGETplus Non-targeting Pool (GE, Cat# D-001810–10-20), ON-TARGETplus Human STUB1 siRNA (short interfering RNA) SMARTpool: ON-TARGETplus STUB1 siRNA (GE, Cat# L007201–00-0020), and ON-TARGETplus Mouse STUB1 siRNA SMARTpool (GE, Cat# L-063143–01-0005). .. Odyssey® Blocking Buffer (PBS) (Li-COR, Cat# 927–40000), QuikChange II XL Site-Directed Mutagenesis Kit (Stratagene, Cat# 200521), Streptavidin MagneSphere® Paramagnetic Particles (Promega, Cat# Z5481), 10× TBS (KD Medical, Cat# RGF-3385), 4× SDS sample buffer (Invitrogen, Cat# NP0008), TRIzol Reagent (Life Technologies, Cat# 15596018), SuperScript III First-Strand Synthesis System for RT-PCR (Life Technologies, Cat# 18080–051), Power SYBR Green PCR Master Mix (Life Technologies, Cat# 4367659), Pierce Silver Stain Kit (Thermo, Cat#24612), papain (Worthington, Cat# ), Basal Medium Eagle (Sigma, Cat# B1522), B27 supplement (Thermo Scientific, Cat# 17504–044), N2 supplement (Thermo Scientific, Cat# 17502–048), glutaMAX-I (Thermo Scientific, Cat# 35050–061), pre-coated coverslips (Corning, Cat# 354087), 0.45% glucose (Sigma, Cat# G8769), and cytosine arabinoside (Sigma, Cat# PHR1787).

    Small Interfering RNA:

    Article Title: The G2385R risk factor for Parkinson’s disease enhances CHIP-dependent intracellular degradation of LRRK2
    Article Snippet: Point mutations were introduced by using the QuikChange II XL Site-Directed Mutagenesis Kit (Stratagene). .. Chemicals and reagents used in the present study were GTP (Sigma, Cat# G8877), protease inhibitor cocktail (Roche, Cat# 04693159001), HALT phosphatase inhibitor cocktail (Thermo Scientific, Cat# 78427), Geldanamycin (Sigma, Cat# G3381), GTP beads (Jena Bioscience, Cat# NU-412–10), EZview Red Protein G Affinity Gel (Sigma, Cat# E3403), EZview Red ANTI-FLAG M2 Affinity Gel (Sigma, Cat# F2426), HaloTag® Biotin Ligand (Promega, Cat# G8281), ON-TARGETplus Non-targeting Pool (GE, Cat# D-001810–10-20), ON-TARGETplus Human STUB1 siRNA (short interfering RNA) SMARTpool: ON-TARGETplus STUB1 siRNA (GE, Cat# L007201–00-0020), and ON-TARGETplus Mouse STUB1 siRNA SMARTpool (GE, Cat# L-063143–01-0005). .. Odyssey® Blocking Buffer (PBS) (Li-COR, Cat# 927–40000), QuikChange II XL Site-Directed Mutagenesis Kit (Stratagene, Cat# 200521), Streptavidin MagneSphere® Paramagnetic Particles (Promega, Cat# Z5481), 10× TBS (KD Medical, Cat# RGF-3385), 4× SDS sample buffer (Invitrogen, Cat# NP0008), TRIzol Reagent (Life Technologies, Cat# 15596018), SuperScript III First-Strand Synthesis System for RT-PCR (Life Technologies, Cat# 18080–051), Power SYBR Green PCR Master Mix (Life Technologies, Cat# 4367659), Pierce Silver Stain Kit (Thermo, Cat#24612), papain (Worthington, Cat# ), Basal Medium Eagle (Sigma, Cat# B1522), B27 supplement (Thermo Scientific, Cat# 17504–044), N2 supplement (Thermo Scientific, Cat# 17502–048), glutaMAX-I (Thermo Scientific, Cat# 35050–061), pre-coated coverslips (Corning, Cat# 354087), 0.45% glucose (Sigma, Cat# G8769), and cytosine arabinoside (Sigma, Cat# PHR1787).

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  • 92
    Jena Bioscience gtpγs
    The catalytic interface is involved in Irga6-Irgb6 and Irga6-Irgm3 interactions . Pull-down of Irgb6 (a) and Irgm3 (b) with recombinant GST-tagged Irga6 protein from IFNγ-stimulated gs3T3 fibroblasts lysate in the presence or absence of guanine nucleotides (0.5 mM GDP, <t>GTPγS</t> or mant-GDP). GST-Irga6 protein was visualised by Ponceau S staining upon blotting (top rows). Irgb6 and Irgm3 were detected with anti-Irgb6 and anti-Irgm3 monoclonal antibodies (bottom rows). A shorter exposure of the lysate input is shown. Dotted lines indicate positions where irrelevant lanes were removed from the image.
    Gtpγs, supplied by Jena Bioscience, 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/gtpγs/product/Jena Bioscience
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gtpγs - by Bioz Stars, 2021-04
    92/100 stars
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    The catalytic interface is involved in Irga6-Irgb6 and Irga6-Irgm3 interactions . Pull-down of Irgb6 (a) and Irgm3 (b) with recombinant GST-tagged Irga6 protein from IFNγ-stimulated gs3T3 fibroblasts lysate in the presence or absence of guanine nucleotides (0.5 mM GDP, GTPγS or mant-GDP). GST-Irga6 protein was visualised by Ponceau S staining upon blotting (top rows). Irgb6 and Irgm3 were detected with anti-Irgb6 and anti-Irgm3 monoclonal antibodies (bottom rows). A shorter exposure of the lysate input is shown. Dotted lines indicate positions where irrelevant lanes were removed from the image.

    Journal: BMC Biology

    Article Title: The activation mechanism of Irga6, an interferon-inducible GTPase contributing to mouse resistance against Toxoplasma gondii

    doi: 10.1186/1741-7007-9-7

    Figure Lengend Snippet: The catalytic interface is involved in Irga6-Irgb6 and Irga6-Irgm3 interactions . Pull-down of Irgb6 (a) and Irgm3 (b) with recombinant GST-tagged Irga6 protein from IFNγ-stimulated gs3T3 fibroblasts lysate in the presence or absence of guanine nucleotides (0.5 mM GDP, GTPγS or mant-GDP). GST-Irga6 protein was visualised by Ponceau S staining upon blotting (top rows). Irgb6 and Irgm3 were detected with anti-Irgb6 and anti-Irgm3 monoclonal antibodies (bottom rows). A shorter exposure of the lysate input is shown. Dotted lines indicate positions where irrelevant lanes were removed from the image.

    Article Snippet: Nucleotides GTP (Carl Roth, Karlsruhe, Germany and Sigma-Aldrich, St.Louis, MO, USA); GDP (Sigma-Aldrich, St.Louis, MO, USA); GTPγS, XTP, 2'deoxy-GTP, mant-GTP, mant-GDP, mant-GTPγS, 2'mant-3'deoxy-GTP, 2'deoxy-3'mant-GTP, mant-XTP and mant-XDP (Jena Bioscience, Jena, Germany); 3'deoxy-GTP (Jena Bioscience, Jena, Germany and Trilink Biotechnologies, San Diego, CA, USA); 2'3'dideoxy-GTP (GE Healthcare, Munich, Germany); α32 P-GTP (GE Healthcare, Munich, Germany, Hartmann Analytic, Braunschweig, Germany and Perkin Elmer, Waltham, MA, USA); γ32 P-3'dGTP (Hartmann Analytic, Braunschweig, Germany)

    Techniques: Recombinant, Staining

    Evidence that s NUCB1 acts as a GDI of Gα i1 . A, we estimated the number of available nucleotide-binding sites on Gα i1 in the absence or presence of Ca 2+ -free s NUCB1 by monitoring the absorbance of Gα i1 -bound BODIPY FL-GTPγS

    Journal: The Journal of Biological Chemistry

    Article Title: Nucleobindin 1 Is a Calcium-regulated Guanine Nucleotide Dissociation Inhibitor of G?i1 *

    doi: 10.1074/jbc.M110.148429

    Figure Lengend Snippet: Evidence that s NUCB1 acts as a GDI of Gα i1 . A, we estimated the number of available nucleotide-binding sites on Gα i1 in the absence or presence of Ca 2+ -free s NUCB1 by monitoring the absorbance of Gα i1 -bound BODIPY FL-GTPγS

    Article Snippet: The fluorescent GTPγS analogue, mant(2′,3′- O -( N -methyl-anthraniloyl)-GTPγS was purchased from Jena Biosciences (Jena, Germany), and BODIPY-FL-GTPγS was purchased from Molecular Probes (Invitrogen).

    Techniques: Binding Assay

    Effect of GTP hydrolysis on binding of IF1 and IF2 to mature 70S IC. ( A ) 30S IC formed in the presence of Bpy-Met-tRNA fMet and GTP (12.5 μM) was rapidly mixed with 50S subunits in the presence or absence of GTPγS (0.25 mM). Time courses of Bpy-Met-tRNA fMet fluorescence changes were monitored. ( B ) Interaction of Bpy-Met-tRNA fMet with IF2 upon binding of the factor to 70S IC was followed by mixing purified 70S ICs (containing Bpy-Met-tRNA fMet ) with IF2 bound to GTPγS, GDPNP, GTP or GDP. Similar experiments were performed using an IF2 variant lacking the C2-domain (ΔC2) in the presence of GTPγS. ( C ) 30S S13 (Alx488) IC formed with IF1 4 (Atto540Q) and GTP (12.5 μM) was mixed with 50S subunits in the presence or absence of GTPγS (0.25 mM). ( D ) The binding of the IF1 to mature 70S IC was followed by mixing non-purified 70S ICs (formed with 30S S13 (Alx488) in the absence of IF1) with IF1 4 (Atto540Q), in the presence of GTP or GTPγS.

    Journal: Nucleic Acids Research

    Article Title: Directional transition from initiation to elongation in bacterial translation

    doi: 10.1093/nar/gkv869

    Figure Lengend Snippet: Effect of GTP hydrolysis on binding of IF1 and IF2 to mature 70S IC. ( A ) 30S IC formed in the presence of Bpy-Met-tRNA fMet and GTP (12.5 μM) was rapidly mixed with 50S subunits in the presence or absence of GTPγS (0.25 mM). Time courses of Bpy-Met-tRNA fMet fluorescence changes were monitored. ( B ) Interaction of Bpy-Met-tRNA fMet with IF2 upon binding of the factor to 70S IC was followed by mixing purified 70S ICs (containing Bpy-Met-tRNA fMet ) with IF2 bound to GTPγS, GDPNP, GTP or GDP. Similar experiments were performed using an IF2 variant lacking the C2-domain (ΔC2) in the presence of GTPγS. ( C ) 30S S13 (Alx488) IC formed with IF1 4 (Atto540Q) and GTP (12.5 μM) was mixed with 50S subunits in the presence or absence of GTPγS (0.25 mM). ( D ) The binding of the IF1 to mature 70S IC was followed by mixing non-purified 70S ICs (formed with 30S S13 (Alx488) in the absence of IF1) with IF1 4 (Atto540Q), in the presence of GTP or GTPγS.

    Article Snippet: GTP, GDP, GTPγS, GDPNP, mant-GTP (2′/3′-O -(N -methyl-anthraniloyl)-guanosine-5′-triphosphate, triethylammonium salt) and mant-GTPγS (2′/3′-O -(N -methyl-anthraniloyl)-guanosine-5′-(γ-thio)-triphosphate, triethylammonium salt), were purchased from Jena Biosciences; Bpy-GTP (guanosine 5′-triphosphate, BODIPY FL 2′-(or-3′)-O -(N -(2-aminoethyl)urethane), trisodium salt) and Bpy-GDP (guanosine 5′-diphosphate, BODIPY FL 2′-(or-3′)-O -(N -(2-aminoethyl)urethane), bis-(triethylammonium) salt) from Life Technologies.

    Techniques: Binding Assay, Fluorescence, Purification, Variant Assay

    Effect of GTP hydrolysis on the kinetics of 70S IC maturation. 30S IC was rapidly mixed with 50S subunits (1 μM) in a stopped-flow ( A – E ) or a quench-flow ( F ) machine and the time courses of indicated reactions were monitored in the presence of GTP (black) or GTPγS (green). An inset in ( F ) shows the extended time window for peptide bond formation. Smooth lines (A–E) show fits obtained by global evaluation of all time courses using numerical integration.

    Journal: Nucleic Acids Research

    Article Title: Directional transition from initiation to elongation in bacterial translation

    doi: 10.1093/nar/gkv869

    Figure Lengend Snippet: Effect of GTP hydrolysis on the kinetics of 70S IC maturation. 30S IC was rapidly mixed with 50S subunits (1 μM) in a stopped-flow ( A – E ) or a quench-flow ( F ) machine and the time courses of indicated reactions were monitored in the presence of GTP (black) or GTPγS (green). An inset in ( F ) shows the extended time window for peptide bond formation. Smooth lines (A–E) show fits obtained by global evaluation of all time courses using numerical integration.

    Article Snippet: GTP, GDP, GTPγS, GDPNP, mant-GTP (2′/3′-O -(N -methyl-anthraniloyl)-guanosine-5′-triphosphate, triethylammonium salt) and mant-GTPγS (2′/3′-O -(N -methyl-anthraniloyl)-guanosine-5′-(γ-thio)-triphosphate, triethylammonium salt), were purchased from Jena Biosciences; Bpy-GTP (guanosine 5′-triphosphate, BODIPY FL 2′-(or-3′)-O -(N -(2-aminoethyl)urethane), trisodium salt) and Bpy-GDP (guanosine 5′-diphosphate, BODIPY FL 2′-(or-3′)-O -(N -(2-aminoethyl)urethane), bis-(triethylammonium) salt) from Life Technologies.

    Techniques: Flow Cytometry