crude histrap column  (GE Healthcare)

 
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    Name:
    HisTrap FF Crude
    Description:

    Catalog Number:
    11000458
    Price:
    None
    Category:
    HisTrap FF Crude histidine tagged protein purification columns
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    Structured Review

    GE Healthcare crude histrap column
    PKA phosphorylates GPKOW at S27 and T316 in vitro . A . Recombinant GPKOW proteins were expressed in bacteria and purified on a <t>HisTrap</t> column followed by an ion exchange column. GPKOW wt (lane 1), GPKOW S27A (lane 2), GPKOW T316A (lane 3) and GPKOW S27A+T316A (lane 4) were all recognized by anti-GPKOW B01 after separation on SDS-PAGE and immunoblotting. One representative immunoblot is shown. B . Purified GPKOW wt (lanes 1 and 2), single- (lanes 3-6) or double-mutated GPKOW (lanes 7 and 8) were incubated with active (+) or heat inactivated (-) PKA Cα1 (7.4 ng) and γ- 32 P-ATP in a reaction buffer. The samples were analyzed by SDS-PAGE and autoradiography. The CBB staining in the lower panel shows the amount of the different proteins. C . Unsaturated images from Syngene G-box and Typhoon 9400 phosphoimager were quantified by Genetools (Syngene) and the statistical analysis was performed with paired students T-test in GraphPad Prism. The asterisk indicates a significant difference between the columns with a p-value

    https://www.bioz.com/result/crude histrap column/product/GE Healthcare
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    crude histrap column - by Bioz Stars, 2021-05
    97/100 stars

    Images

    1) Product Images from "G-patch domain and KOW motifs-containing protein, GPKOW; a nuclear RNA-binding protein regulated by protein kinase A"

    Article Title: G-patch domain and KOW motifs-containing protein, GPKOW; a nuclear RNA-binding protein regulated by protein kinase A

    Journal: Journal of Molecular Signaling

    doi: 10.1186/1750-2187-6-10

    PKA phosphorylates GPKOW at S27 and T316 in vitro . A . Recombinant GPKOW proteins were expressed in bacteria and purified on a HisTrap column followed by an ion exchange column. GPKOW wt (lane 1), GPKOW S27A (lane 2), GPKOW T316A (lane 3) and GPKOW S27A+T316A (lane 4) were all recognized by anti-GPKOW B01 after separation on SDS-PAGE and immunoblotting. One representative immunoblot is shown. B . Purified GPKOW wt (lanes 1 and 2), single- (lanes 3-6) or double-mutated GPKOW (lanes 7 and 8) were incubated with active (+) or heat inactivated (-) PKA Cα1 (7.4 ng) and γ- 32 P-ATP in a reaction buffer. The samples were analyzed by SDS-PAGE and autoradiography. The CBB staining in the lower panel shows the amount of the different proteins. C . Unsaturated images from Syngene G-box and Typhoon 9400 phosphoimager were quantified by Genetools (Syngene) and the statistical analysis was performed with paired students T-test in GraphPad Prism. The asterisk indicates a significant difference between the columns with a p-value
    Figure Legend Snippet: PKA phosphorylates GPKOW at S27 and T316 in vitro . A . Recombinant GPKOW proteins were expressed in bacteria and purified on a HisTrap column followed by an ion exchange column. GPKOW wt (lane 1), GPKOW S27A (lane 2), GPKOW T316A (lane 3) and GPKOW S27A+T316A (lane 4) were all recognized by anti-GPKOW B01 after separation on SDS-PAGE and immunoblotting. One representative immunoblot is shown. B . Purified GPKOW wt (lanes 1 and 2), single- (lanes 3-6) or double-mutated GPKOW (lanes 7 and 8) were incubated with active (+) or heat inactivated (-) PKA Cα1 (7.4 ng) and γ- 32 P-ATP in a reaction buffer. The samples were analyzed by SDS-PAGE and autoradiography. The CBB staining in the lower panel shows the amount of the different proteins. C . Unsaturated images from Syngene G-box and Typhoon 9400 phosphoimager were quantified by Genetools (Syngene) and the statistical analysis was performed with paired students T-test in GraphPad Prism. The asterisk indicates a significant difference between the columns with a p-value

    Techniques Used: In Vitro, Recombinant, Purification, SDS Page, Incubation, Autoradiography, Staining

    Related Articles

    other:

    Article Title: Enhancement of Soluble Expression and Biochemical Characterization of Two Epoxide Hydrolases from Bacillus
    Article Snippet: HisTrap FF Crude 1 mL columns were purchased from GE Healthcare (Boston, MA, USA).

    Article Title: The Mammalian Cytosolic Type 2 (R)-β-hydroxybutyrate Dehydrogenase (BDH2) is 4-oxo-L-proline Reductase (EC 1.1.1.104)
    Article Snippet: Q-Sepharose FF resin, HiScreen Blue FF, Superdex 200 16/60 HiLoad, and HisTrap FF crude columns were obtained from GE Healthcare Bio-Sciences (Uppsala, Sweden).

    Binding Assay:

    Article Title: Construction of an artificial biosynthetic pathway for hyperextended archaeal membrane lipids in the bacterium Escherichia coli
    Article Snippet: After an additional 24 h of incubation, the cells were harvested and disrupted by sonication in a HisTrap binding buffer that contained 20 mM potassium phosphate, pH 7.4, 0.5 M NaCl and 10 mM imidazole. .. The supernatant fraction was loaded into a HisTrap crude FF column (GE Healthcare, USA), which had been equilibrated with HisTrap binding buffer. ..

    Protease Inhibitor:

    Article Title: G-patch domain and KOW motifs-containing protein, GPKOW; a nuclear RNA-binding protein regulated by protein kinase A
    Article Snippet: The bacteria were pelleted and resuspended in 500 ml TB-medium and grown to optic density between 0.5-0.7 at 600 nm at 37°C and 150 rpm, before protein expression was induced by 0.25 mM IPTG and further grown for 4 hours at 25°C at 150 rpm. .. The bacteria were lysed in Buffer A (50 mM sodium dihydrogen phosphate pH 7.5, 0.2 M NaCl, 5 mM Imidazole, 1 × EDTA free protease inhibitor cocktail (Roche 04693132001) and 0.4 mM betamercaptoetanol) and purified on a crude HisTrap column (GE Healthcare 11-0004-58) using Buffer A and Buffer B (50 mM sodium dihydrogen phosphate pH 7.5, 0.2 M NaCl, 500 mM Imidazole, 1 × EDTA free protease inhibitor cocktail (Roche 04693132001) and 0.4 mM betamercaptoetanol). .. Fractions with the highest optical density at 280 nm were pooled, dialyzed to Buffer C (20 mM Tris-HCl, pH 8) and further purified by ion exchange chromatography (Resource Q, GE Healthcare 520348) at pH 8 with buffer C and buffer D (20 mM Tris-HCl pH 8, 1 M NaCl).

    Purification:

    Article Title: G-patch domain and KOW motifs-containing protein, GPKOW; a nuclear RNA-binding protein regulated by protein kinase A
    Article Snippet: The bacteria were pelleted and resuspended in 500 ml TB-medium and grown to optic density between 0.5-0.7 at 600 nm at 37°C and 150 rpm, before protein expression was induced by 0.25 mM IPTG and further grown for 4 hours at 25°C at 150 rpm. .. The bacteria were lysed in Buffer A (50 mM sodium dihydrogen phosphate pH 7.5, 0.2 M NaCl, 5 mM Imidazole, 1 × EDTA free protease inhibitor cocktail (Roche 04693132001) and 0.4 mM betamercaptoetanol) and purified on a crude HisTrap column (GE Healthcare 11-0004-58) using Buffer A and Buffer B (50 mM sodium dihydrogen phosphate pH 7.5, 0.2 M NaCl, 500 mM Imidazole, 1 × EDTA free protease inhibitor cocktail (Roche 04693132001) and 0.4 mM betamercaptoetanol). .. Fractions with the highest optical density at 280 nm were pooled, dialyzed to Buffer C (20 mM Tris-HCl, pH 8) and further purified by ion exchange chromatography (Resource Q, GE Healthcare 520348) at pH 8 with buffer C and buffer D (20 mM Tris-HCl pH 8, 1 M NaCl).

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    GE Healthcare crude histrap column
    PKA phosphorylates GPKOW at S27 and T316 in vitro . A . Recombinant GPKOW proteins were expressed in bacteria and purified on a <t>HisTrap</t> column followed by an ion exchange column. GPKOW wt (lane 1), GPKOW S27A (lane 2), GPKOW T316A (lane 3) and GPKOW S27A+T316A (lane 4) were all recognized by anti-GPKOW B01 after separation on SDS-PAGE and immunoblotting. One representative immunoblot is shown. B . Purified GPKOW wt (lanes 1 and 2), single- (lanes 3-6) or double-mutated GPKOW (lanes 7 and 8) were incubated with active (+) or heat inactivated (-) PKA Cα1 (7.4 ng) and γ- 32 P-ATP in a reaction buffer. The samples were analyzed by SDS-PAGE and autoradiography. The CBB staining in the lower panel shows the amount of the different proteins. C . Unsaturated images from Syngene G-box and Typhoon 9400 phosphoimager were quantified by Genetools (Syngene) and the statistical analysis was performed with paired students T-test in GraphPad Prism. The asterisk indicates a significant difference between the columns with a p-value
    Crude Histrap Column, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/crude histrap column/product/GE Healthcare
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    crude histrap column - by Bioz Stars, 2021-05
    97/100 stars
      Buy from Supplier

    97
    GE Healthcare histrap ff crude column
    Purification and oligomeric state of CdFic SE/AA . ( a ) Coomassie-stained 4–15% gradient SDS–PAGE of purified CdFic SE/AA . Lane M , molecular-weight marker (labelled in kDa). Lane 1, elution from the <t>HisTrap</t> affinity column, 1.8 µg
    Histrap Ff Crude Column, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/histrap ff crude column/product/GE Healthcare
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    histrap ff crude column - by Bioz Stars, 2021-05
    97/100 stars
      Buy from Supplier

    Image Search Results


    PKA phosphorylates GPKOW at S27 and T316 in vitro . A . Recombinant GPKOW proteins were expressed in bacteria and purified on a HisTrap column followed by an ion exchange column. GPKOW wt (lane 1), GPKOW S27A (lane 2), GPKOW T316A (lane 3) and GPKOW S27A+T316A (lane 4) were all recognized by anti-GPKOW B01 after separation on SDS-PAGE and immunoblotting. One representative immunoblot is shown. B . Purified GPKOW wt (lanes 1 and 2), single- (lanes 3-6) or double-mutated GPKOW (lanes 7 and 8) were incubated with active (+) or heat inactivated (-) PKA Cα1 (7.4 ng) and γ- 32 P-ATP in a reaction buffer. The samples were analyzed by SDS-PAGE and autoradiography. The CBB staining in the lower panel shows the amount of the different proteins. C . Unsaturated images from Syngene G-box and Typhoon 9400 phosphoimager were quantified by Genetools (Syngene) and the statistical analysis was performed with paired students T-test in GraphPad Prism. The asterisk indicates a significant difference between the columns with a p-value

    Journal: Journal of Molecular Signaling

    Article Title: G-patch domain and KOW motifs-containing protein, GPKOW; a nuclear RNA-binding protein regulated by protein kinase A

    doi: 10.1186/1750-2187-6-10

    Figure Lengend Snippet: PKA phosphorylates GPKOW at S27 and T316 in vitro . A . Recombinant GPKOW proteins were expressed in bacteria and purified on a HisTrap column followed by an ion exchange column. GPKOW wt (lane 1), GPKOW S27A (lane 2), GPKOW T316A (lane 3) and GPKOW S27A+T316A (lane 4) were all recognized by anti-GPKOW B01 after separation on SDS-PAGE and immunoblotting. One representative immunoblot is shown. B . Purified GPKOW wt (lanes 1 and 2), single- (lanes 3-6) or double-mutated GPKOW (lanes 7 and 8) were incubated with active (+) or heat inactivated (-) PKA Cα1 (7.4 ng) and γ- 32 P-ATP in a reaction buffer. The samples were analyzed by SDS-PAGE and autoradiography. The CBB staining in the lower panel shows the amount of the different proteins. C . Unsaturated images from Syngene G-box and Typhoon 9400 phosphoimager were quantified by Genetools (Syngene) and the statistical analysis was performed with paired students T-test in GraphPad Prism. The asterisk indicates a significant difference between the columns with a p-value

    Article Snippet: The bacteria were lysed in Buffer A (50 mM sodium dihydrogen phosphate pH 7.5, 0.2 M NaCl, 5 mM Imidazole, 1 × EDTA free protease inhibitor cocktail (Roche 04693132001) and 0.4 mM betamercaptoetanol) and purified on a crude HisTrap column (GE Healthcare 11-0004-58) using Buffer A and Buffer B (50 mM sodium dihydrogen phosphate pH 7.5, 0.2 M NaCl, 500 mM Imidazole, 1 × EDTA free protease inhibitor cocktail (Roche 04693132001) and 0.4 mM betamercaptoetanol).

    Techniques: In Vitro, Recombinant, Purification, SDS Page, Incubation, Autoradiography, Staining

    A , expression and purification of recombinant proteins. Proteins were expressed in E. coli BL21(DE3) strain. All proteins were expressed in fusion with the His 6 tag. Expressed recombinant proteins were purified using HisTrap FF crude columns (GE Healthcare).

    Journal: The Journal of Biological Chemistry

    Article Title: Biochemical Characterization and Substrate Specificity of Autophagin-2 from the Parasite Trypanosoma cruzi *

    doi: 10.1074/jbc.M115.687764

    Figure Lengend Snippet: A , expression and purification of recombinant proteins. Proteins were expressed in E. coli BL21(DE3) strain. All proteins were expressed in fusion with the His 6 tag. Expressed recombinant proteins were purified using HisTrap FF crude columns (GE Healthcare).

    Article Snippet: All proteins were expressed as fusion protein with a His6 tag and were allowed to bind to HisTrap FF crude columns (GE Healthcare) and eluted with buffer containing 250 m m imidazole.

    Techniques: Expressing, Purification, Recombinant

    Purification and oligomeric state of CdFic SE/AA . ( a ) Coomassie-stained 4–15% gradient SDS–PAGE of purified CdFic SE/AA . Lane M , molecular-weight marker (labelled in kDa). Lane 1, elution from the HisTrap affinity column, 1.8 µg

    Journal: Acta Crystallographica. Section F, Structural Biology Communications

    Article Title: Protein expression, characterization, crystallization and preliminary X-ray crystallographic analysis of a Fic protein from Clostridium difficile

    doi: 10.1107/S2053230X1400987X

    Figure Lengend Snippet: Purification and oligomeric state of CdFic SE/AA . ( a ) Coomassie-stained 4–15% gradient SDS–PAGE of purified CdFic SE/AA . Lane M , molecular-weight marker (labelled in kDa). Lane 1, elution from the HisTrap affinity column, 1.8 µg

    Article Snippet: The resulting supernatant was filtered with a 0.45 µm syringe filter before application onto a pre-packed 1 ml HisTrap FF Crude column (GE Healthcare) charged with Ni2+ and washed with 20 column volumes of 50 m M Tris pH 7.4, 300 m M NaCl, 5 m M β-mercaptoethanol, 5%( v / v ) glycerol, 20 m M imidazole.

    Techniques: Purification, Staining, SDS Page, Molecular Weight, Marker, Affinity Column

    Purified trKMO. (A) Chromatogram produced during NiNTA affinity purification of truncate KMO. (B) SDS-PAGE protein gel stained with simplyblue showing HisTrap purification of truncated human KMO protein, (a) Cell-free extract prior to affinity chromatography, (b) flowthrough proteins which did not bind to the column, (c) first column wash, (d) second column wash, (e) third column wash, (f) pure truncated human KMO protein (44 kDa) from the fractions under the chromatogram. (C) Cartoon illustrating the trKMO construct. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Journal: Protein Expression and Purification

    Article Title: Bacterial expression of human kynurenine 3-monooxygenase: Solubility, activity, purification

    doi: 10.1016/j.pep.2013.11.015

    Figure Lengend Snippet: Purified trKMO. (A) Chromatogram produced during NiNTA affinity purification of truncate KMO. (B) SDS-PAGE protein gel stained with simplyblue showing HisTrap purification of truncated human KMO protein, (a) Cell-free extract prior to affinity chromatography, (b) flowthrough proteins which did not bind to the column, (c) first column wash, (d) second column wash, (e) third column wash, (f) pure truncated human KMO protein (44 kDa) from the fractions under the chromatogram. (C) Cartoon illustrating the trKMO construct. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: The soluble supernatant fraction was purified on a 5 ml HisTrap ff crude column (GE Healthcare, 17-5286-01) using the AKTA protein purification system (GE Healthcare).

    Techniques: Purification, Produced, Affinity Purification, SDS Page, Staining, Affinity Chromatography, Construct