nucleotide analogue maba adp  (Jena Bioscience)


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  • 94
    Name:
    8 4 Amino butyl amino ADP MANT
    Description:

    Catalog Number:
    NU-893-MNT
    Price:
    921.5
    Category:
    Nucleotides Nucleosides
    Size:
    200 µl
    Buy from Supplier


    Structured Review

    Jena Bioscience nucleotide analogue maba adp
    MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of <t>MABA-ADP</t> and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P

    https://www.bioz.com/result/nucleotide analogue maba adp/product/Jena Bioscience
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nucleotide analogue maba adp - by Bioz Stars, 2021-06
    94/100 stars

    Images

    1) Product Images from "MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP"

    Article Title: MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP

    Journal: Nature Communications

    doi: 10.1038/s41467-019-08450-4

    MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of MABA-ADP and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P
    Figure Legend Snippet: MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of MABA-ADP and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P

    Techniques Used: Fluorescence, Isolation, Mutagenesis

    2) Product Images from "MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP"

    Article Title: MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP

    Journal: Nature Communications

    doi: 10.1038/s41467-019-08450-4

    MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of MABA-ADP and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P
    Figure Legend Snippet: MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of MABA-ADP and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P

    Techniques Used: Fluorescence, Isolation, Mutagenesis

    3) Product Images from "AMPylation targets the rate-limiting step of BiP’s ATPase cycle for its functional inactivation"

    Article Title: AMPylation targets the rate-limiting step of BiP’s ATPase cycle for its functional inactivation

    Journal: eLife

    doi: 10.7554/eLife.29428

    Grp170 stimulates MABA-ADP release from BiP. ( A ) Coomassie (CBB)-stained SDS-PAGE gels of bacterially expressed and purified human Grp170 protein carrying an N-terminal His6-tag. A sample of the protein preparation was applied to SDS-PAGE (left) and the full-length protein (FL) as well as co-purified faster migrating species (D and 1–4) are indicated. The latter contain mainly Grp170-derived peptides as identified by mass spectrometry analysis ( Supplementary file 1 ). A Ni-NTA affinity pull-down (right) under native conditions or upon denaturation with 6 M guanidine hydrochloride (GdnHCl) supports their identity as N-terminal proteolytic fragments of Grp170 (carrying a His6-tag) and indicates only minor contamination of the preparation with other proteins. ( B ) Measurement of nucleotide release from BiP or DnaK in presence of 2 mM calcium. Unmodified wildtype BiP (or DnaK) protein (2.5 µM) was incubated with the fluorescent ADP derivative MABA-ADP (at 2.5 µM) and the dissociation of the formed complexes was measured over time upon 1:1 dilution with a solution containing 3 mM ATP without or with Grp170 (at 0.9 µM or 1.4 µM) as depicted on the scheme. MABA-ADP fluorescence was detected by excitation at 360 nm and recording the emission signal at 440 nm. The Coomassie-stained SDS-PAGE gel on the right shows purified DnaK used in this assay. ( C ) Representative measurement of MABA-ADP release from unmodified or AMPylated BiP (BiP-AMP) as in ‘B’ upon dilution into an ATP-containing solution without or with 1.4 µM Grp170. The raw fluorescence traces were fitted to a single exponential function (grey curves) to calculate the dissociation rate constants presented in Figure 6C .
    Figure Legend Snippet: Grp170 stimulates MABA-ADP release from BiP. ( A ) Coomassie (CBB)-stained SDS-PAGE gels of bacterially expressed and purified human Grp170 protein carrying an N-terminal His6-tag. A sample of the protein preparation was applied to SDS-PAGE (left) and the full-length protein (FL) as well as co-purified faster migrating species (D and 1–4) are indicated. The latter contain mainly Grp170-derived peptides as identified by mass spectrometry analysis ( Supplementary file 1 ). A Ni-NTA affinity pull-down (right) under native conditions or upon denaturation with 6 M guanidine hydrochloride (GdnHCl) supports their identity as N-terminal proteolytic fragments of Grp170 (carrying a His6-tag) and indicates only minor contamination of the preparation with other proteins. ( B ) Measurement of nucleotide release from BiP or DnaK in presence of 2 mM calcium. Unmodified wildtype BiP (or DnaK) protein (2.5 µM) was incubated with the fluorescent ADP derivative MABA-ADP (at 2.5 µM) and the dissociation of the formed complexes was measured over time upon 1:1 dilution with a solution containing 3 mM ATP without or with Grp170 (at 0.9 µM or 1.4 µM) as depicted on the scheme. MABA-ADP fluorescence was detected by excitation at 360 nm and recording the emission signal at 440 nm. The Coomassie-stained SDS-PAGE gel on the right shows purified DnaK used in this assay. ( C ) Representative measurement of MABA-ADP release from unmodified or AMPylated BiP (BiP-AMP) as in ‘B’ upon dilution into an ATP-containing solution without or with 1.4 µM Grp170. The raw fluorescence traces were fitted to a single exponential function (grey curves) to calculate the dissociation rate constants presented in Figure 6C .

    Techniques Used: Staining, SDS Page, Purification, Derivative Assay, Mass Spectrometry, Incubation, Fluorescence

    Related Articles

    Concentration Assay:

    Article Title: The cytoprotective protein MANF promotes neuronal survival independently from its role as a GRP78 cofactor
    Article Snippet: MABA–ADP release assay Assaying the release of fluorescent ADP-analog MABA–ADP was done essentially as has been published before ( ). .. Briefly, 5 μM GRP78 was mixed with MABA–ADP (NU-893-MNT, Jena Bioscience) in equal volume and equimolar concentration and incubated for 3 h at 30 °C while protected from light. .. Nucleotide exchange solutions contained 250 μM ATP (R0441, Thermo Fisher Scientific) and proteins as indicated.

    Incubation:

    Article Title: The cytoprotective protein MANF promotes neuronal survival independently from its role as a GRP78 cofactor
    Article Snippet: MABA–ADP release assay Assaying the release of fluorescent ADP-analog MABA–ADP was done essentially as has been published before ( ). .. Briefly, 5 μM GRP78 was mixed with MABA–ADP (NU-893-MNT, Jena Bioscience) in equal volume and equimolar concentration and incubated for 3 h at 30 °C while protected from light. .. Nucleotide exchange solutions contained 250 μM ATP (R0441, Thermo Fisher Scientific) and proteins as indicated.

    Article Title: Simple methods for the 3′ biotinylation of RNA
    Article Snippet: Chemically synthesized RNA N22 ( , biomers.net) was 5′-labeled with [γ-32 P]ATP and T4 polynucleotide kinase (NEB). .. Up to 2 µM RNA was incubated with 1–5 µM tCid1 in 100 mM potassium acetate, 20 mM Tris-HCl (pH 8.0), 2 mM magnesium acetate, 0.05% NP-40 at 38°C for the indicated time in the presence of 0.5 mM ATP or N6-ATP analog (N6-[(6-amino)hexyl]-amino-ATP-biotin) or 8-ATP analog (8-[(6-amino)hexyl]-amino-ATP-Biotin; both from Jena Bioscience, Germany). ..

    Article Title: AMPylation targets the rate-limiting step of BiP’s ATPase cycle for its functional inactivation
    Article Snippet: The signals were quantified using Image J64 and the ADP values were normalized to the total radioactive signal (ADP + ATP) for each time point and fitted with a single exponential function using the GraphPad Prism 6 software. .. ADP release experiments Non-AMPylated or AMPylated BiP proteins at 1 µM were incubated with 1 µM of the fluorescent ADP analog MABA-ADP (8-[(4-Amino)butyl]-amino-ADP - MANT; Jena Bioscience, Jena, Germany, cat. no. NU-893-MNT) ( ) in HKM buffer for 2 hr at 30°C. .. Afterwards, the samples were mixed at 25°C in a 1:1 (v/v) ratio with HKM containing 2 mM ATP using a stopped-flow reaction analyzer (SX.18MV; Applied Photophysics, Leatherhead, United Kingdom).

    Fluorescence:

    Article Title: MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP
    Article Snippet: Molecular graphics were generated using UCSF Chimera and PyMol (The PyMOL Molecular Graphics System, Version 1.3 Schrödinger, LLC). .. Fluorescence measurement Nucleotide release kinetics of the fluorescent nucleotide analogue MABA-ADP (Jena Bioscience, cat. # NU-893-MNT) and nucleotide binding of MABA-ATP (Jena Bioscience, cat. # NU-806-MNT) were performed in a Perkin Elmer LS55 fluorescence spectrometer instrument (excitation 360 nm, emission 420 nm). .. These nucleotide analogues carry a fluorescent MANT group attached to the ribose, which shows a significant increase of fluorescence upon Hsp70 binding .

    Article Title: MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP
    Article Snippet: Molecular graphics were generated using UCSF Chimera and PyMol (The PyMOL Molecular Graphics System, Version 1.3 Schrödinger, LLC). .. Nucleotide release kinetics of the fluorescent nucleotide analogue MABA-ADP (Jena Bioscience, cat. # NU-893-MNT) and nucleotide binding of MABA-ATP (Jena Bioscience, cat. # NU-806-MNT) were performed in a Perkin Elmer LS55 fluorescence spectrometer instrument (excitation 360 nm, emission 420 nm). .. These nucleotide analogues carry a fluorescent MANT group attached to the ribose, which shows a significant increase of fluorescence upon Hsp70 binding .

    Article Title: Calcium depletion challenges endoplasmic reticulum proteostasis by destabilising BiP-substrate complexes
    Article Snippet: Standard curves from serial dilutions of KH2 PO4 served as a reference to calculate the specific ATPase activity. .. Nucleotide binding assaysNucleotide release and binding measurements were performed using the fluorescent nucleotide analogues MABA-ADP (Jena Bioscience, NU-893-MNT) and MABA-ATP (Jena Bioscience, NU-806-MNT) carrying a MANT moiety whose fluorescence increases upon binding to Hsp70s ( ). .. The measurements were started by adding 55 μl of solution A to 5 μl of solution B (specified below) in a quartz cuvette (Hellma Analytics) and fluorescence (excitation 360 nm, emission 420 nm) was detected over time with a fluorescence spectrometer (Perkin Elmer LS55) at 26°C.

    Article Title: Calcium depletion challenges endoplasmic reticulum proteostasis by destabilising BiP-substrate complexes
    Article Snippet: Standard curves from serial dilutions of KH2 PO4 served as a reference to calculate the specific ATPase activity. .. Nucleotide- binding assaysNucleotide release and binding measurements were performed using the fluorescent nucleotide analogues MABA-ADP (Jena Bioscience, cat. # NU-893-MNT) and MABA-ATP (Jena Bioscience, cat. # NU-806-MNT) carrying a MANT moiety whose fluorescence increases upon binding to Hsp70s ( ). .. The measurements were started by manually adding 55 µl of solution A to 5 µl of solution B (specified below) in a quartz cuvette (Hellma Analytics) and fluorescence (excitation 360 nm, emission 420 nm) was detected over time with a fluorescence spectrometer (Perkin Elmer LS55) at 26°C.

    Binding Assay:

    Article Title: MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP
    Article Snippet: Molecular graphics were generated using UCSF Chimera and PyMol (The PyMOL Molecular Graphics System, Version 1.3 Schrödinger, LLC). .. Fluorescence measurement Nucleotide release kinetics of the fluorescent nucleotide analogue MABA-ADP (Jena Bioscience, cat. # NU-893-MNT) and nucleotide binding of MABA-ATP (Jena Bioscience, cat. # NU-806-MNT) were performed in a Perkin Elmer LS55 fluorescence spectrometer instrument (excitation 360 nm, emission 420 nm). .. These nucleotide analogues carry a fluorescent MANT group attached to the ribose, which shows a significant increase of fluorescence upon Hsp70 binding .

    Article Title: MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP
    Article Snippet: Molecular graphics were generated using UCSF Chimera and PyMol (The PyMOL Molecular Graphics System, Version 1.3 Schrödinger, LLC). .. Nucleotide release kinetics of the fluorescent nucleotide analogue MABA-ADP (Jena Bioscience, cat. # NU-893-MNT) and nucleotide binding of MABA-ATP (Jena Bioscience, cat. # NU-806-MNT) were performed in a Perkin Elmer LS55 fluorescence spectrometer instrument (excitation 360 nm, emission 420 nm). .. These nucleotide analogues carry a fluorescent MANT group attached to the ribose, which shows a significant increase of fluorescence upon Hsp70 binding .

    Article Title: Calcium depletion challenges endoplasmic reticulum proteostasis by destabilising BiP-substrate complexes
    Article Snippet: Standard curves from serial dilutions of KH2 PO4 served as a reference to calculate the specific ATPase activity. .. Nucleotide binding assaysNucleotide release and binding measurements were performed using the fluorescent nucleotide analogues MABA-ADP (Jena Bioscience, NU-893-MNT) and MABA-ATP (Jena Bioscience, NU-806-MNT) carrying a MANT moiety whose fluorescence increases upon binding to Hsp70s ( ). .. The measurements were started by adding 55 μl of solution A to 5 μl of solution B (specified below) in a quartz cuvette (Hellma Analytics) and fluorescence (excitation 360 nm, emission 420 nm) was detected over time with a fluorescence spectrometer (Perkin Elmer LS55) at 26°C.

    Article Title: Calcium depletion challenges endoplasmic reticulum proteostasis by destabilising BiP-substrate complexes
    Article Snippet: Standard curves from serial dilutions of KH2 PO4 served as a reference to calculate the specific ATPase activity. .. Nucleotide- binding assaysNucleotide release and binding measurements were performed using the fluorescent nucleotide analogues MABA-ADP (Jena Bioscience, cat. # NU-893-MNT) and MABA-ATP (Jena Bioscience, cat. # NU-806-MNT) carrying a MANT moiety whose fluorescence increases upon binding to Hsp70s ( ). .. The measurements were started by manually adding 55 µl of solution A to 5 µl of solution B (specified below) in a quartz cuvette (Hellma Analytics) and fluorescence (excitation 360 nm, emission 420 nm) was detected over time with a fluorescence spectrometer (Perkin Elmer LS55) at 26°C.

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  • 94
    Jena Bioscience nucleotide analogue maba adp
    MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of <t>MABA-ADP</t> and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P
    Nucleotide Analogue Maba Adp, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nucleotide analogue maba adp/product/Jena Bioscience
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nucleotide analogue maba adp - by Bioz Stars, 2021-06
    94/100 stars
      Buy from Supplier

    Image Search Results


    MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of MABA-ADP and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P

    Journal: Nature Communications

    Article Title: MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP

    doi: 10.1038/s41467-019-08450-4

    Figure Lengend Snippet: MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of MABA-ADP and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P

    Article Snippet: Fluorescence measurement Nucleotide release kinetics of the fluorescent nucleotide analogue MABA-ADP (Jena Bioscience, cat. # NU-893-MNT) and nucleotide binding of MABA-ATP (Jena Bioscience, cat. # NU-806-MNT) were performed in a Perkin Elmer LS55 fluorescence spectrometer instrument (excitation 360 nm, emission 420 nm).

    Techniques: Fluorescence, Isolation, Mutagenesis

    MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of MABA-ADP and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P

    Journal: Nature Communications

    Article Title: MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP

    doi: 10.1038/s41467-019-08450-4

    Figure Lengend Snippet: MANF inhibits nucleotide exchange and ATP-induced substrate release from BiP. a Representative plot of fluorescence against time of pre-formed complexes of MABA-ADP and BiP NBD (1.25 µM) challenged at t = 0 with buffer solution containing ATP (125 µM) and MANF or its SAP domain (200 µM). b Bar diagram of MABA-ADP release rates from BiP NBD in the presence of MANF, the isolated SAP or SAPLIP domain or the indicated mutant forms of MANF. Bars represent mean values ± SD from three to five independent experiments (**** P

    Article Snippet: Nucleotide release kinetics of the fluorescent nucleotide analogue MABA-ADP (Jena Bioscience, cat. # NU-893-MNT) and nucleotide binding of MABA-ATP (Jena Bioscience, cat. # NU-806-MNT) were performed in a Perkin Elmer LS55 fluorescence spectrometer instrument (excitation 360 nm, emission 420 nm).

    Techniques: Fluorescence, Isolation, Mutagenesis

    Grp170 stimulates MABA-ADP release from BiP. ( A ) Coomassie (CBB)-stained SDS-PAGE gels of bacterially expressed and purified human Grp170 protein carrying an N-terminal His6-tag. A sample of the protein preparation was applied to SDS-PAGE (left) and the full-length protein (FL) as well as co-purified faster migrating species (D and 1–4) are indicated. The latter contain mainly Grp170-derived peptides as identified by mass spectrometry analysis ( Supplementary file 1 ). A Ni-NTA affinity pull-down (right) under native conditions or upon denaturation with 6 M guanidine hydrochloride (GdnHCl) supports their identity as N-terminal proteolytic fragments of Grp170 (carrying a His6-tag) and indicates only minor contamination of the preparation with other proteins. ( B ) Measurement of nucleotide release from BiP or DnaK in presence of 2 mM calcium. Unmodified wildtype BiP (or DnaK) protein (2.5 µM) was incubated with the fluorescent ADP derivative MABA-ADP (at 2.5 µM) and the dissociation of the formed complexes was measured over time upon 1:1 dilution with a solution containing 3 mM ATP without or with Grp170 (at 0.9 µM or 1.4 µM) as depicted on the scheme. MABA-ADP fluorescence was detected by excitation at 360 nm and recording the emission signal at 440 nm. The Coomassie-stained SDS-PAGE gel on the right shows purified DnaK used in this assay. ( C ) Representative measurement of MABA-ADP release from unmodified or AMPylated BiP (BiP-AMP) as in ‘B’ upon dilution into an ATP-containing solution without or with 1.4 µM Grp170. The raw fluorescence traces were fitted to a single exponential function (grey curves) to calculate the dissociation rate constants presented in Figure 6C .

    Journal: eLife

    Article Title: AMPylation targets the rate-limiting step of BiP’s ATPase cycle for its functional inactivation

    doi: 10.7554/eLife.29428

    Figure Lengend Snippet: Grp170 stimulates MABA-ADP release from BiP. ( A ) Coomassie (CBB)-stained SDS-PAGE gels of bacterially expressed and purified human Grp170 protein carrying an N-terminal His6-tag. A sample of the protein preparation was applied to SDS-PAGE (left) and the full-length protein (FL) as well as co-purified faster migrating species (D and 1–4) are indicated. The latter contain mainly Grp170-derived peptides as identified by mass spectrometry analysis ( Supplementary file 1 ). A Ni-NTA affinity pull-down (right) under native conditions or upon denaturation with 6 M guanidine hydrochloride (GdnHCl) supports their identity as N-terminal proteolytic fragments of Grp170 (carrying a His6-tag) and indicates only minor contamination of the preparation with other proteins. ( B ) Measurement of nucleotide release from BiP or DnaK in presence of 2 mM calcium. Unmodified wildtype BiP (or DnaK) protein (2.5 µM) was incubated with the fluorescent ADP derivative MABA-ADP (at 2.5 µM) and the dissociation of the formed complexes was measured over time upon 1:1 dilution with a solution containing 3 mM ATP without or with Grp170 (at 0.9 µM or 1.4 µM) as depicted on the scheme. MABA-ADP fluorescence was detected by excitation at 360 nm and recording the emission signal at 440 nm. The Coomassie-stained SDS-PAGE gel on the right shows purified DnaK used in this assay. ( C ) Representative measurement of MABA-ADP release from unmodified or AMPylated BiP (BiP-AMP) as in ‘B’ upon dilution into an ATP-containing solution without or with 1.4 µM Grp170. The raw fluorescence traces were fitted to a single exponential function (grey curves) to calculate the dissociation rate constants presented in Figure 6C .

    Article Snippet: ADP release experiments Non-AMPylated or AMPylated BiP proteins at 1 µM were incubated with 1 µM of the fluorescent ADP analog MABA-ADP (8-[(4-Amino)butyl]-amino-ADP - MANT; Jena Bioscience, Jena, Germany, cat. no. NU-893-MNT) ( ) in HKM buffer for 2 hr at 30°C.

    Techniques: Staining, SDS Page, Purification, Derivative Assay, Mass Spectrometry, Incubation, Fluorescence