penta his hrp conjugate kit  (Qiagen)


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    Penta His HRP Conjugate Kit
    Description:
    For sensitive direct detection of His tagged proteins Kit contents Qiagen PentaHis HRP Conjugate Kit His tagged Protein HHHH Epitopes HRP Conjugated Enzyme Mouse IgG1 Antibody Isotype Secondary Antibody Required OPD Substrates for Assay Procedure 4C1N Substrate for Blot Detection High sensitivity and High speed Detection Ideal for Western Dot and Colony Blotting LISA and Other Immunoassay Procedures Includes 125L Penta His HRP Conjugate 5g Blocking Reagent 50mL Blocking Reagent Buffer 10x Concentrate Benefits High sensitivity and high speed detection of His tagged proteins No need for secondary antibody Complete kit with optimized blocking r
    Catalog Number:
    34460
    Price:
    516
    Category:
    Penta His HRP Conjugate Kit
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    Structured Review

    Qiagen penta his hrp conjugate kit
    Penta His HRP Conjugate Kit
    For sensitive direct detection of His tagged proteins Kit contents Qiagen PentaHis HRP Conjugate Kit His tagged Protein HHHH Epitopes HRP Conjugated Enzyme Mouse IgG1 Antibody Isotype Secondary Antibody Required OPD Substrates for Assay Procedure 4C1N Substrate for Blot Detection High sensitivity and High speed Detection Ideal for Western Dot and Colony Blotting LISA and Other Immunoassay Procedures Includes 125L Penta His HRP Conjugate 5g Blocking Reagent 50mL Blocking Reagent Buffer 10x Concentrate Benefits High sensitivity and high speed detection of His tagged proteins No need for secondary antibody Complete kit with optimized blocking r
    https://www.bioz.com/result/penta his hrp conjugate kit/product/Qiagen
    Average 99 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    penta his hrp conjugate kit - by Bioz Stars, 2020-04
    99/100 stars

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    1) Product Images from "Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins"

    Article Title: Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins

    Journal: Microbial Cell Factories

    doi: 10.1186/1475-2859-11-66

    Overproduction of different secretory proteins in B. subtilis. ( a ) Whole cell extracts, ( b ) medium fractions, ( c ) membrane fractions, ( d ) cytoplasmic fractions. Left panels show SDS-PAGE gels; right panels ( b , c and d ) show immunodetection of the 6his-tagged proteins using Penta-His HRP conjugate antibodies (Qiagen). Asterisks indicate protein bands corresponding to the overproduced proteins. Calculated molecular masses of proteins, with and without signal peptide, in kDa: XynA, 32.3 (preprotein), 20.4 (matured); NprE, 56.5 (preprotein), 53.9 (matured); Bla (TEM-1 β-lactamase), 32.3 (preprotein), 29.7 (matured); Usp45, 48.2 (preprotein), 45.5 (matured); MntA, 33.4 (preprotein), 32.4 (matured); YcdH, 36.5 (preprotein), 34.3 (matured); LmrA, 66.2; XylP, 55.3.
    Figure Legend Snippet: Overproduction of different secretory proteins in B. subtilis. ( a ) Whole cell extracts, ( b ) medium fractions, ( c ) membrane fractions, ( d ) cytoplasmic fractions. Left panels show SDS-PAGE gels; right panels ( b , c and d ) show immunodetection of the 6his-tagged proteins using Penta-His HRP conjugate antibodies (Qiagen). Asterisks indicate protein bands corresponding to the overproduced proteins. Calculated molecular masses of proteins, with and without signal peptide, in kDa: XynA, 32.3 (preprotein), 20.4 (matured); NprE, 56.5 (preprotein), 53.9 (matured); Bla (TEM-1 β-lactamase), 32.3 (preprotein), 29.7 (matured); Usp45, 48.2 (preprotein), 45.5 (matured); MntA, 33.4 (preprotein), 32.4 (matured); YcdH, 36.5 (preprotein), 34.3 (matured); LmrA, 66.2; XylP, 55.3.

    Techniques Used: SDS Page, Immunodetection, Transmission Electron Microscopy

    Related Articles

    Nucleic Acid Electrophoresis:

    Article Title: Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses
    Article Snippet: His-tagged CPs were separated by SDS-polyacrylamide gel electrophoresis (PAGE) with NuPAGE Novex 12% Bis-tris Gels (Thermo Fisher Scientific). .. Both purified recombinant CPs were detected by immunoblotting using a previously described method [ ] with a modification that involved the use of penta-His HRP conjugate kit (Qiagen, Venlo, The Netherlands).

    Article Title: Structure-Activity Relationship of Gelatinase Biosynthesis-Activating Pheromone of Enterococcus faecalis ▿
    Article Snippet: The membrane fraction was separated by ultracentrifugation at 125,000 × g for 90 min and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by Western blot analysis using anti-histidine tag antibody to detect FsrC-His6 . .. Western blot analysis was performed using the Penta-His HRP conjugate kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer's protocol.

    Centrifugation:

    Article Title: Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins
    Article Snippet: 1 OD unit of a culture was collected by centrifugation, resuspended in 150 μl of buffer con taining 10 mM Tris–HCl pH 8.1, 20% sucrose, 10 mM EDTA, 50 mM NaCl and 2 mg/ml lysozyme, and incubated at 37°C for 30 min. An equal volume of 2x SDS-PAGE sample buffer (100 mM Tris–HCl pH 6.8, 4% SDS, 1% DTT, 20% glycerol, 0.05% bromophenol blue) was added and the samples were boiled for 5 min. .. The immunodetection of His-tagged proteins was performed using the Penta-His HRP Conjugate Kit (Qiagen) and ECL detection reagents (Amersham).

    Incubation:

    Article Title: A Novel Virus-Like Particle Based Vaccine Platform Displaying the Placental Malaria Antigen VAR2CSA
    Article Snippet: Conjugation and purification of mSA-VAR2CSA to L1-Avi VLPs For production of the VLP-VAR2CSA vaccine, biotinylated HPV16 Avi-L1 VLPs were mixed with 3x molar excess of mSA-VAR2CSA and the sample was incubated at 4°C for 18–24 hours with gentle shaking. .. Post ultracentrifugation fractions were analyzed by SDS-PAGE or blotted onto a nitrocellulose membrane (GE-Healthcare RPN203E) for detection of L1 or mSA-VAR2CSA (via a 6xhistidine tag) with CamVir-1 (AbD Serotec, Bio-Rad, 7135–2804) or Penta His HRP (QIAGEN, 34460) respectively.

    Article Title: Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins
    Article Snippet: The extracellular proteins present in the medium were precipitated by adding 200 μl of ice-cold 100% TCA to 1.8 ml of medium and incubation on ice for 1 hour. .. The immunodetection of His-tagged proteins was performed using the Penta-His HRP Conjugate Kit (Qiagen) and ECL detection reagents (Amersham).

    Article Title: FmvB: A Francisella tularensis Magnesium-Responsive Outer Membrane Protein that Plays a Role in Virulence
    Article Snippet: .. Immunoblotting For detection of FopA (outer membrane control), SecY (inner membrane control), and 6x-histidine fusion tagged FmvB in sucrose gradient fractions, samples were diluted in SDS-PAGE loading buffer and boiled for 10 min. For comparison, whole cell lysates of ΔfmvB and ΔfmvB (pFNLTP6-gro-fmvB -6xHis) were prepared by suspension in SDS-PAGE loading buffer and boiling for 10 min. Molecular mass standards (Precision Plus protein all blue prestained protein standards; BioRad Laboratories) and protein samples were separated by SDS-PAGE, transferred to nitrocellulose, blots were incubated overnight in blot block (0.1% (vol/vol) Tween 20 and 2% (wt/vol) bovine serum albumin in PBS) at 4°C, and immunoblotting was performed using rat polyclonal antiserum specific for either F . tularensis FopA or F . tularensis SecY [ ] or the Penta-His HRP conjugate antibody (Qiagen). .. FopA antiserum was diluted 1:10,000 in blot block, SecY antiserum was diluted 1:5000 in blot block, and Penta-His HRP conjugate antibody was diluted 1:10,000 in block.

    Expressing:

    Article Title: Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses
    Article Snippet: Recombinant proteins were expressed by culturing Escherichia coli strain Rosetta (DE3) (Merck Millipore, Billerica, MA, USA) harboring each expression vector for 3 h at 30 °C in the presence of 1 mM isopropyl-β-d (–)-thiogalactopyranoside. .. Both purified recombinant CPs were detected by immunoblotting using a previously described method [ ] with a modification that involved the use of penta-His HRP conjugate kit (Qiagen, Venlo, The Netherlands).

    Article Title: Structure-Activity Relationship of Gelatinase Biosynthesis-Activating Pheromone of Enterococcus faecalis ▿
    Article Snippet: The culture was maintained for 3 h for the expression of FsrC-His6 . .. Western blot analysis was performed using the Penta-His HRP conjugate kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer's protocol.

    Article Title: FmvB: A Francisella tularensis Magnesium-Responsive Outer Membrane Protein that Plays a Role in Virulence
    Article Snippet: Immunoblotting For detection of FopA (outer membrane control), SecY (inner membrane control), and 6x-histidine fusion tagged FmvB in sucrose gradient fractions, samples were diluted in SDS-PAGE loading buffer and boiled for 10 min. For comparison, whole cell lysates of ΔfmvB and ΔfmvB (pFNLTP6-gro-fmvB -6xHis) were prepared by suspension in SDS-PAGE loading buffer and boiling for 10 min. Molecular mass standards (Precision Plus protein all blue prestained protein standards; BioRad Laboratories) and protein samples were separated by SDS-PAGE, transferred to nitrocellulose, blots were incubated overnight in blot block (0.1% (vol/vol) Tween 20 and 2% (wt/vol) bovine serum albumin in PBS) at 4°C, and immunoblotting was performed using rat polyclonal antiserum specific for either F . tularensis FopA or F . tularensis SecY [ ] or the Penta-His HRP conjugate antibody (Qiagen). .. To compare LPS expression levels between LVS and ΔfmvB grown in magnesium-limiting medium, bacteria were harvested by centrifugation, adjusted to OD600 0.8, suspended in SDS-PAGE loading buffer, and boiled for 10 min.

    Modification:

    Article Title: Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses
    Article Snippet: .. Both purified recombinant CPs were detected by immunoblotting using a previously described method [ ] with a modification that involved the use of penta-His HRP conjugate kit (Qiagen, Venlo, The Netherlands). .. Anti-BSSV rabbit polyclonal antibody (Agdia Inc., Elkhart, IN, USA) and goat anti-rabbit HRP conjugate (Bio-Rad, Hercules, CA, USA) were used for the specific detection of BSSV-CP.

    Conjugation Assay:

    Article Title: A Novel Virus-Like Particle Based Vaccine Platform Displaying the Placental Malaria Antigen VAR2CSA
    Article Snippet: Paragraph title: Conjugation and purification of mSA-VAR2CSA to L1-Avi VLPs ... Post ultracentrifugation fractions were analyzed by SDS-PAGE or blotted onto a nitrocellulose membrane (GE-Healthcare RPN203E) for detection of L1 or mSA-VAR2CSA (via a 6xhistidine tag) with CamVir-1 (AbD Serotec, Bio-Rad, 7135–2804) or Penta His HRP (QIAGEN, 34460) respectively.

    Isolation:

    Article Title: Inhibition of Staphylococcus aureus by Lysostaphin-Expressing Lactobacillus plantarum WCFS1 in a Modified Genital Tract Secretion Medium ▿ WCFS1 in a Modified Genital Tract Secretion Medium ▿ †
    Article Snippet: Proteins from spent medium isolated from overnight cultures (15 to 18 h) of L. plantarum WCFS1 grown in buffered (pH 7.0) mGTS were resolved by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes (Amersham Hybond-P; GE Healthcare Bio-Sciences, Piscataway, NJ) using the semidry method as recommended by the manufacturer (Bio-Rad, Hercules, CA). .. The penta-His antibody–horseradish peroxidase (HRP) conjugate kit (Qiagen, Valencia, CA) and a chemiluminescence kit (GE Healthcare) were used as recommended by their respective manufacturers for detection of the His6 -tagged lysostaphin.

    Size-exclusion Chromatography:

    Article Title: Characterization of Atg38 and NRBF2, a fifth subunit of the autophagic Vps34/PIK3C3 complex
    Article Snippet: The cells were treated using a cell wall loosening procedure (2 M LiAc on ice for 3 min followed by 0.4 M NaOH on ice for 3 min), and disrupted in 50 mM Tris (pH 8.0), 1% SDS, 8 M urea (VWR, 28877), 2 mM DTT, 5 mM EDTA with glass beads at 3,000 rpm for 30 sec with PowerLyzer homogenizer (MO BIO Laboratories, 13155, Carlsbad, California, USA) at 4°C. .. For detecting the His-tagged human NRBF2 MIT domain, Penta-His HRP conjugate Kit (Qiagen, 34660) was used with BenchMark (Invitrogen, 10747012) as a marker.

    Blocking Assay:

    Article Title: FmvB: A Francisella tularensis Magnesium-Responsive Outer Membrane Protein that Plays a Role in Virulence
    Article Snippet: .. FopA antiserum was diluted 1:10,000 in blot block, SecY antiserum was diluted 1:5000 in blot block, and Penta-His HRP conjugate antibody was diluted 1:10,000 in block. .. Blots were incubated with primary antisera for 2 h at room temperature, washed four times for 5 min each with blot washing buffer (0.05% [vol/vol] Tween 20 in PBS), FopA and SecY blots were incubated with goat anti-rat IgG (H+L)-HRP secondary antibody (Jackson ImmunoResearch Laboratories) for 1 h at room temperature, washed 3 times for 5 min each with blot washing buffer, washed one time for 5 min in PBS, and blots were developed by either colorimetric detection reagent (FopA; 0.06% [wt/vol] 4-chloro-1-naphthol (Sigma), 10.4% [vol/vol] methanol, and 0.13% [vol/vol] hydrogen peroxide in 200 mM NaCl and 50 mM Tris base) or chemiluminescent detection reagent (SecY and Penta-His HRP; SuperSignal Chemiluminescent Substrate; Thermo Scientific).

    Article Title: FmvB: A Francisella tularensis Magnesium-Responsive Outer Membrane Protein that Plays a Role in Virulence
    Article Snippet: .. Immunoblotting For detection of FopA (outer membrane control), SecY (inner membrane control), and 6x-histidine fusion tagged FmvB in sucrose gradient fractions, samples were diluted in SDS-PAGE loading buffer and boiled for 10 min. For comparison, whole cell lysates of ΔfmvB and ΔfmvB (pFNLTP6-gro-fmvB -6xHis) were prepared by suspension in SDS-PAGE loading buffer and boiling for 10 min. Molecular mass standards (Precision Plus protein all blue prestained protein standards; BioRad Laboratories) and protein samples were separated by SDS-PAGE, transferred to nitrocellulose, blots were incubated overnight in blot block (0.1% (vol/vol) Tween 20 and 2% (wt/vol) bovine serum albumin in PBS) at 4°C, and immunoblotting was performed using rat polyclonal antiserum specific for either F . tularensis FopA or F . tularensis SecY [ ] or the Penta-His HRP conjugate antibody (Qiagen). .. FopA antiserum was diluted 1:10,000 in blot block, SecY antiserum was diluted 1:5000 in blot block, and Penta-His HRP conjugate antibody was diluted 1:10,000 in block.

    Immunodetection:

    Article Title: Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins
    Article Snippet: .. The immunodetection of His-tagged proteins was performed using the Penta-His HRP Conjugate Kit (Qiagen) and ECL detection reagents (Amersham). ..

    Concentration Assay:

    Article Title: Structure-Activity Relationship of Gelatinase Biosynthesis-Activating Pheromone of Enterococcus faecalis ▿
    Article Snippet: After the culture reached an OD600 of 0.5, nisin A (Sigma-Aldrich, Inc., St. Louis, MO) dissolved in water and adjusted to pH 3.0 with HCl was added to achieve a final concentration of 10 ng ml−1 for the induction of the fsrC -His6 gene. .. Western blot analysis was performed using the Penta-His HRP conjugate kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer's protocol.

    Article Title: A Novel Virus-Like Particle Based Vaccine Platform Displaying the Placental Malaria Antigen VAR2CSA
    Article Snippet: The VLP-VAR2CSA vaccine was dialyzed in PBS (0.32 M NaCl, 0.02% PS80) and the relative concentration of VLP-bound mSA-VAR2CSA was estimated by SDS-PAGE using a BSA standard dilution range. .. Post ultracentrifugation fractions were analyzed by SDS-PAGE or blotted onto a nitrocellulose membrane (GE-Healthcare RPN203E) for detection of L1 or mSA-VAR2CSA (via a 6xhistidine tag) with CamVir-1 (AbD Serotec, Bio-Rad, 7135–2804) or Penta His HRP (QIAGEN, 34460) respectively.

    Purification:

    Article Title: Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses
    Article Snippet: .. Both purified recombinant CPs were detected by immunoblotting using a previously described method [ ] with a modification that involved the use of penta-His HRP conjugate kit (Qiagen, Venlo, The Netherlands). .. Anti-BSSV rabbit polyclonal antibody (Agdia Inc., Elkhart, IN, USA) and goat anti-rabbit HRP conjugate (Bio-Rad, Hercules, CA, USA) were used for the specific detection of BSSV-CP.

    Article Title: A Novel Virus-Like Particle Based Vaccine Platform Displaying the Placental Malaria Antigen VAR2CSA
    Article Snippet: Paragraph title: Conjugation and purification of mSA-VAR2CSA to L1-Avi VLPs ... Post ultracentrifugation fractions were analyzed by SDS-PAGE or blotted onto a nitrocellulose membrane (GE-Healthcare RPN203E) for detection of L1 or mSA-VAR2CSA (via a 6xhistidine tag) with CamVir-1 (AbD Serotec, Bio-Rad, 7135–2804) or Penta His HRP (QIAGEN, 34460) respectively.

    Polymerase Chain Reaction:

    Article Title: Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses
    Article Snippet: The PCR products were introduced into a penta-His tagged protein expression vector pET28a (Merck Millipore, Darmstadt, Germany) using the following restriction enzymes: NheI and XhoI for BSSV-CP, and NdeI and XhoI for PIW-CP. .. Both purified recombinant CPs were detected by immunoblotting using a previously described method [ ] with a modification that involved the use of penta-His HRP conjugate kit (Qiagen, Venlo, The Netherlands).

    Recombinant:

    Article Title: Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses
    Article Snippet: .. Both purified recombinant CPs were detected by immunoblotting using a previously described method [ ] with a modification that involved the use of penta-His HRP conjugate kit (Qiagen, Venlo, The Netherlands). .. Anti-BSSV rabbit polyclonal antibody (Agdia Inc., Elkhart, IN, USA) and goat anti-rabbit HRP conjugate (Bio-Rad, Hercules, CA, USA) were used for the specific detection of BSSV-CP.

    Article Title: Optimization of EnBase Fed-Batch Cultivation to Improve Soluble Fraction Ratio of α-Luffin Ribosome Inactivating Protein
    Article Snippet: .. Confirmation of Expressed Recombinant Protein Cellular pellet was run on 12% SDS-PAGE and western-blotting was performed using mouse anti-His antibody, Penta-His HRP Conjugate, (QIAGEN, USA). .. Protein Purification Using Ni-NTA Column E. coli cells were resuspended in lysis buffer (2-5 mL.g-1 ), 50 mM NaH2 PO4 (pH 8.0), containing 300 mM NaCl and 20 mM imidazole in order to increase the purity of recombinant protein and to minimize binding of undesired contaminating proteins.

    Article Title: Optimization of EnBase Fed-Batch Cultivation to Improve Soluble Fraction Ratio of α-Luffin Ribosome Inactivating Protein
    Article Snippet: Paragraph title: 3.7. Confirmation of Expressed Recombinant Protein ... Cellular pellet was run on 12% SDS-PAGE and western-blotting was performed using mouse anti-His antibody, Penta-His HRP Conjugate, (QIAGEN, USA).

    Plasmid Preparation:

    Article Title: Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses
    Article Snippet: Recombinant proteins were expressed by culturing Escherichia coli strain Rosetta (DE3) (Merck Millipore, Billerica, MA, USA) harboring each expression vector for 3 h at 30 °C in the presence of 1 mM isopropyl-β-d (–)-thiogalactopyranoside. .. Both purified recombinant CPs were detected by immunoblotting using a previously described method [ ] with a modification that involved the use of penta-His HRP conjugate kit (Qiagen, Venlo, The Netherlands).

    Polyacrylamide Gel Electrophoresis:

    Article Title: Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses
    Article Snippet: His-tagged CPs were separated by SDS-polyacrylamide gel electrophoresis (PAGE) with NuPAGE Novex 12% Bis-tris Gels (Thermo Fisher Scientific). .. Both purified recombinant CPs were detected by immunoblotting using a previously described method [ ] with a modification that involved the use of penta-His HRP conjugate kit (Qiagen, Venlo, The Netherlands).

    Marker:

    Article Title: Characterization of Atg38 and NRBF2, a fifth subunit of the autophagic Vps34/PIK3C3 complex
    Article Snippet: .. For detecting the His-tagged human NRBF2 MIT domain, Penta-His HRP conjugate Kit (Qiagen, 34660) was used with BenchMark (Invitrogen, 10747012) as a marker. .. Subcellular fractionation Subcellular fractionation was performed using a modified version of a described procedure.

    Western Blot:

    Article Title: A widespread bacterial type VI secretion effector superfamily identified using a heuristic approach
    Article Snippet: .. The α-His5 Western blots were performed using the Penta-His HRP Conjugate Kit according to manufacturer’s instructions (Qiagen). ..

    Article Title: Structure-Activity Relationship of Gelatinase Biosynthesis-Activating Pheromone of Enterococcus faecalis ▿
    Article Snippet: .. Western blot analysis was performed using the Penta-His HRP conjugate kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer's protocol. .. After the induction of recombinant L. lactis cells, the cells were harvested and washed with 50 mM Tris-HCl (pH 7.8), and the cell concentration was adjusted to an OD600 of 10.

    Article Title: Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins
    Article Snippet: Paragraph title: SDS-PAGE and Western blotting ... The immunodetection of His-tagged proteins was performed using the Penta-His HRP Conjugate Kit (Qiagen) and ECL detection reagents (Amersham).

    Article Title: Optimization of EnBase Fed-Batch Cultivation to Improve Soluble Fraction Ratio of α-Luffin Ribosome Inactivating Protein
    Article Snippet: .. Confirmation of Expressed Recombinant Protein Cellular pellet was run on 12% SDS-PAGE and western-blotting was performed using mouse anti-His antibody, Penta-His HRP Conjugate, (QIAGEN, USA). .. Protein Purification Using Ni-NTA Column E. coli cells were resuspended in lysis buffer (2-5 mL.g-1 ), 50 mM NaH2 PO4 (pH 8.0), containing 300 mM NaCl and 20 mM imidazole in order to increase the purity of recombinant protein and to minimize binding of undesired contaminating proteins.

    Article Title: Inhibition of Staphylococcus aureus by Lysostaphin-Expressing Lactobacillus plantarum WCFS1 in a Modified Genital Tract Secretion Medium ▿ WCFS1 in a Modified Genital Tract Secretion Medium ▿ †
    Article Snippet: Paragraph title: Western analysis. ... The penta-His antibody–horseradish peroxidase (HRP) conjugate kit (Qiagen, Valencia, CA) and a chemiluminescence kit (GE Healthcare) were used as recommended by their respective manufacturers for detection of the His6 -tagged lysostaphin.

    Article Title: Characterization of Atg38 and NRBF2, a fifth subunit of the autophagic Vps34/PIK3C3 complex
    Article Snippet: Paragraph title: Western blotting ... For detecting the His-tagged human NRBF2 MIT domain, Penta-His HRP conjugate Kit (Qiagen, 34660) was used with BenchMark (Invitrogen, 10747012) as a marker.

    Article Title: Optimization of EnBase Fed-Batch Cultivation to Improve Soluble Fraction Ratio of α-Luffin Ribosome Inactivating Protein
    Article Snippet: .. Cellular pellet was run on 12% SDS-PAGE and western-blotting was performed using mouse anti-His antibody, Penta-His HRP Conjugate, (QIAGEN, USA). .. E. coli cells were resuspended in lysis buffer (2-5 mL.g-1 ), 50 mM NaH2 PO4 (pH 8.0), containing 300 mM NaCl and 20 mM imidazole in order to increase the purity of recombinant protein and to minimize binding of undesired contaminating proteins.

    Over Expression:

    Article Title: Structure-Activity Relationship of Gelatinase Biosynthesis-Activating Pheromone of Enterococcus faecalis ▿
    Article Snippet: Paragraph title: Overexpression of FsrC on Lactococcus lactis . ... Western blot analysis was performed using the Penta-His HRP conjugate kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer's protocol.

    other:

    Article Title: A photo-cross-linking approach to monitor folding and assembly of newly synthesized proteins in a living cell
    Article Snippet: Penta-His HRP conjugate was purchased from Qiagen (Hilden, Germany).

    SDS Page:

    Article Title: A Novel Virus-Like Particle Based Vaccine Platform Displaying the Placental Malaria Antigen VAR2CSA
    Article Snippet: .. Post ultracentrifugation fractions were analyzed by SDS-PAGE or blotted onto a nitrocellulose membrane (GE-Healthcare RPN203E) for detection of L1 or mSA-VAR2CSA (via a 6xhistidine tag) with CamVir-1 (AbD Serotec, Bio-Rad, 7135–2804) or Penta His HRP (QIAGEN, 34460) respectively. .. Immunization of mice Female C57BL/6 mice (Taconic, Denmark) were immunized by intramuscular injection with 5 μg VLP-coupled mSA-VAR2CSA, soluble mSA-VAR2CSA or soluble naked VAR2CSA on day 0 with no adjuvant.

    Article Title: Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins
    Article Snippet: Paragraph title: SDS-PAGE and Western blotting ... The immunodetection of His-tagged proteins was performed using the Penta-His HRP Conjugate Kit (Qiagen) and ECL detection reagents (Amersham).

    Article Title: Optimization of EnBase Fed-Batch Cultivation to Improve Soluble Fraction Ratio of α-Luffin Ribosome Inactivating Protein
    Article Snippet: .. Confirmation of Expressed Recombinant Protein Cellular pellet was run on 12% SDS-PAGE and western-blotting was performed using mouse anti-His antibody, Penta-His HRP Conjugate, (QIAGEN, USA). .. Protein Purification Using Ni-NTA Column E. coli cells were resuspended in lysis buffer (2-5 mL.g-1 ), 50 mM NaH2 PO4 (pH 8.0), containing 300 mM NaCl and 20 mM imidazole in order to increase the purity of recombinant protein and to minimize binding of undesired contaminating proteins.

    Article Title: Inhibition of Staphylococcus aureus by Lysostaphin-Expressing Lactobacillus plantarum WCFS1 in a Modified Genital Tract Secretion Medium ▿ WCFS1 in a Modified Genital Tract Secretion Medium ▿ †
    Article Snippet: Proteins from spent medium isolated from overnight cultures (15 to 18 h) of L. plantarum WCFS1 grown in buffered (pH 7.0) mGTS were resolved by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes (Amersham Hybond-P; GE Healthcare Bio-Sciences, Piscataway, NJ) using the semidry method as recommended by the manufacturer (Bio-Rad, Hercules, CA). .. The penta-His antibody–horseradish peroxidase (HRP) conjugate kit (Qiagen, Valencia, CA) and a chemiluminescence kit (GE Healthcare) were used as recommended by their respective manufacturers for detection of the His6 -tagged lysostaphin.

    Article Title: FmvB: A Francisella tularensis Magnesium-Responsive Outer Membrane Protein that Plays a Role in Virulence
    Article Snippet: .. Immunoblotting For detection of FopA (outer membrane control), SecY (inner membrane control), and 6x-histidine fusion tagged FmvB in sucrose gradient fractions, samples were diluted in SDS-PAGE loading buffer and boiled for 10 min. For comparison, whole cell lysates of ΔfmvB and ΔfmvB (pFNLTP6-gro-fmvB -6xHis) were prepared by suspension in SDS-PAGE loading buffer and boiling for 10 min. Molecular mass standards (Precision Plus protein all blue prestained protein standards; BioRad Laboratories) and protein samples were separated by SDS-PAGE, transferred to nitrocellulose, blots were incubated overnight in blot block (0.1% (vol/vol) Tween 20 and 2% (wt/vol) bovine serum albumin in PBS) at 4°C, and immunoblotting was performed using rat polyclonal antiserum specific for either F . tularensis FopA or F . tularensis SecY [ ] or the Penta-His HRP conjugate antibody (Qiagen). .. FopA antiserum was diluted 1:10,000 in blot block, SecY antiserum was diluted 1:5000 in blot block, and Penta-His HRP conjugate antibody was diluted 1:10,000 in block.

    Article Title: Optimization of EnBase Fed-Batch Cultivation to Improve Soluble Fraction Ratio of α-Luffin Ribosome Inactivating Protein
    Article Snippet: .. Cellular pellet was run on 12% SDS-PAGE and western-blotting was performed using mouse anti-His antibody, Penta-His HRP Conjugate, (QIAGEN, USA). .. E. coli cells were resuspended in lysis buffer (2-5 mL.g-1 ), 50 mM NaH2 PO4 (pH 8.0), containing 300 mM NaCl and 20 mM imidazole in order to increase the purity of recombinant protein and to minimize binding of undesired contaminating proteins.

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  • 99
    Qiagen penta his hrp conjugate kit
    Overproduction of different secretory proteins in B. subtilis. ( a ) Whole cell extracts, ( b ) medium fractions, ( c ) membrane fractions, ( d ) cytoplasmic fractions. Left panels show SDS-PAGE gels; right panels ( b , c and d ) show <t>immunodetection</t> of the 6his-tagged proteins using Penta-His <t>HRP</t> conjugate antibodies (Qiagen). Asterisks indicate protein bands corresponding to the overproduced proteins. Calculated molecular masses of proteins, with and without signal peptide, in kDa: XynA, 32.3 (preprotein), 20.4 (matured); NprE, 56.5 (preprotein), 53.9 (matured); Bla (TEM-1 β-lactamase), 32.3 (preprotein), 29.7 (matured); Usp45, 48.2 (preprotein), 45.5 (matured); MntA, 33.4 (preprotein), 32.4 (matured); YcdH, 36.5 (preprotein), 34.3 (matured); LmrA, 66.2; XylP, 55.3.
    Penta His Hrp Conjugate Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Overproduction of different secretory proteins in B. subtilis. ( a ) Whole cell extracts, ( b ) medium fractions, ( c ) membrane fractions, ( d ) cytoplasmic fractions. Left panels show SDS-PAGE gels; right panels ( b , c and d ) show immunodetection of the 6his-tagged proteins using Penta-His HRP conjugate antibodies (Qiagen). Asterisks indicate protein bands corresponding to the overproduced proteins. Calculated molecular masses of proteins, with and without signal peptide, in kDa: XynA, 32.3 (preprotein), 20.4 (matured); NprE, 56.5 (preprotein), 53.9 (matured); Bla (TEM-1 β-lactamase), 32.3 (preprotein), 29.7 (matured); Usp45, 48.2 (preprotein), 45.5 (matured); MntA, 33.4 (preprotein), 32.4 (matured); YcdH, 36.5 (preprotein), 34.3 (matured); LmrA, 66.2; XylP, 55.3.

    Journal: Microbial Cell Factories

    Article Title: Comparative transcriptional analysis of Bacillus subtilis cells overproducing either secreted proteins, lipoproteins or membrane proteins

    doi: 10.1186/1475-2859-11-66

    Figure Lengend Snippet: Overproduction of different secretory proteins in B. subtilis. ( a ) Whole cell extracts, ( b ) medium fractions, ( c ) membrane fractions, ( d ) cytoplasmic fractions. Left panels show SDS-PAGE gels; right panels ( b , c and d ) show immunodetection of the 6his-tagged proteins using Penta-His HRP conjugate antibodies (Qiagen). Asterisks indicate protein bands corresponding to the overproduced proteins. Calculated molecular masses of proteins, with and without signal peptide, in kDa: XynA, 32.3 (preprotein), 20.4 (matured); NprE, 56.5 (preprotein), 53.9 (matured); Bla (TEM-1 β-lactamase), 32.3 (preprotein), 29.7 (matured); Usp45, 48.2 (preprotein), 45.5 (matured); MntA, 33.4 (preprotein), 32.4 (matured); YcdH, 36.5 (preprotein), 34.3 (matured); LmrA, 66.2; XylP, 55.3.

    Article Snippet: The immunodetection of His-tagged proteins was performed using the Penta-His HRP Conjugate Kit (Qiagen) and ECL detection reagents (Amersham).

    Techniques: SDS Page, Immunodetection, Transmission Electron Microscopy