ni nta chromatography  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Name:
    Ni NTA Magnetic Beads
    Description:

    Catalog Number:
    S1423
    Price:
    899
    Category:
    Beads Racks Columns
    Applications:
    Proteomics & Glycomics
    Size:
    5 ml
    Buy from Supplier


    Structured Review

    New England Biolabs ni nta chromatography
    Ni NTA Magnetic Beads

    https://www.bioz.com/result/ni nta chromatography/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ni nta chromatography - by Bioz Stars, 2021-08
    95/100 stars

    Images

    1) Product Images from "GAPDH controls extracellular vesicle biogenesis and enhances therapeutic potential of EVs in silencing the Huntingtin gene in mice via siRNA delivery"

    Article Title: GAPDH controls extracellular vesicle biogenesis and enhances therapeutic potential of EVs in silencing the Huntingtin gene in mice via siRNA delivery

    Journal: bioRxiv

    doi: 10.1101/2020.01.09.899880

    G58T-mediated loading of siRNA on EVs: SDS-PAGE of the G58T-based proteins purified by Ni-NTA chromatography. Gel is stained with Coomassie brilliant blue dye. (b) Gel shift assay of G58T EVs (upper) and G58TF EVs (lower), reflecting binding of siRNA to EVs. 20pmoles of siRNA were incubated with the given number of EVs. Bands trapped near the wells represent bound siRNA. siRNA alone was used as negative control. Based on the gel shift assay, around 550 siRNAs bind to each G58T EV and 714 siRNAs to each G58TF EV. Higher binding of siRNA to G58TF is due to the arginine-rich FHV peptide. (c) RNase A protection assay of G58TF EVs bound to 50 pmoles of siRNA. Bands in the agarose gel represent siRNA isolated from G58TF EVs after treatment with RNase A (0.2 mg/ml). G58TF EVs bound to siRNA were incubated with RNase A for 6 h at 37°C. (d) Representative confocal microscopy images of N2a cells after 4h of treatment with G58TF EVs carrying FAM-labelled siRNA (green). Cells were treated with lysotracker red dye (red dots) to label late-endosomes. Inset in the merged figure represents magnified image of N2a cells showing colocalization of siRNA with lysotracker dye (yellow spots). Images were captured using 60X objective lens of Olympus confocal microscope FV1000. (e) GAPDH gene silencing by G58T EVs at mRNA level in N2a cells. Different amounts of GAPDH siRNA bound to G58T EVs were added to cells. Level of mRNA was quantified after 48h of treatment by probe-based real time PCR. Data were analysed using linear regression analysis and ΔΔCt method. Level of mRNA in control was assigned as 1 and used to determine the relative level of GAPDH mRNA in the treated cells. Results are shown as mean ± s.d, n= 3, ***p
    Figure Legend Snippet: G58T-mediated loading of siRNA on EVs: SDS-PAGE of the G58T-based proteins purified by Ni-NTA chromatography. Gel is stained with Coomassie brilliant blue dye. (b) Gel shift assay of G58T EVs (upper) and G58TF EVs (lower), reflecting binding of siRNA to EVs. 20pmoles of siRNA were incubated with the given number of EVs. Bands trapped near the wells represent bound siRNA. siRNA alone was used as negative control. Based on the gel shift assay, around 550 siRNAs bind to each G58T EV and 714 siRNAs to each G58TF EV. Higher binding of siRNA to G58TF is due to the arginine-rich FHV peptide. (c) RNase A protection assay of G58TF EVs bound to 50 pmoles of siRNA. Bands in the agarose gel represent siRNA isolated from G58TF EVs after treatment with RNase A (0.2 mg/ml). G58TF EVs bound to siRNA were incubated with RNase A for 6 h at 37°C. (d) Representative confocal microscopy images of N2a cells after 4h of treatment with G58TF EVs carrying FAM-labelled siRNA (green). Cells were treated with lysotracker red dye (red dots) to label late-endosomes. Inset in the merged figure represents magnified image of N2a cells showing colocalization of siRNA with lysotracker dye (yellow spots). Images were captured using 60X objective lens of Olympus confocal microscope FV1000. (e) GAPDH gene silencing by G58T EVs at mRNA level in N2a cells. Different amounts of GAPDH siRNA bound to G58T EVs were added to cells. Level of mRNA was quantified after 48h of treatment by probe-based real time PCR. Data were analysed using linear regression analysis and ΔΔCt method. Level of mRNA in control was assigned as 1 and used to determine the relative level of GAPDH mRNA in the treated cells. Results are shown as mean ± s.d, n= 3, ***p

    Techniques Used: SDS Page, Purification, Chromatography, Staining, Electrophoretic Mobility Shift Assay, Binding Assay, Incubation, Negative Control, Agarose Gel Electrophoresis, Isolation, Confocal Microscopy, Microscopy, Real-time Polymerase Chain Reaction

    Surface binding of GAPDH leads to aggregation of EVs: (a) Western blot showing exogenous binding of GAPDH to HEK293T EVs. Increasing concentrations of histidine-(His6) and Flag-tagged GAPDH (lane 3,4 and 5) were incubated with a fixed number of EVs (see details in methods). Endogenous GAPDH (Endo. GAPDH) present naturally on the EV surface is shown in the top blot along with the exogenous GAPDH (Exo. GAPDH). Alix and CD81 are EV protein markers used as a positive control. Calnexin (bottom blot) was used to demonstrate the purity of EV samples. In this blot, lane 2, 3 and 4 represent EVs, and lane 5 represents cell lysate. Representative blots (n > 3). (b) UV-absorbance spectrum of EVs after passing through gel-filtration column. Increase in the absorbance of EVs+GAPDH peak indicates binding of GAPDH protein. Purified EVs were used for incubation with either GAPDH (upper chromatogram) or BSA protein (lower chromatogram). The first peak (at ∼10 ml elution) represents EVs and the second peak (around 20 ml elution) represents unbound proteins. Representative graphs (n > 3). (c) NTA profile showing the size distribution of purified HEK293T EVs after incubation with either GAPDH or BSA proteins respectively. Binding of GAPDH to the EVs, shifts their size. Inset is the scatter plot representing size (mean) of EVs (red; EVs+GAPDH, black; EVs+BSA). Data shown as mean ± s.d, n=9, ***p
    Figure Legend Snippet: Surface binding of GAPDH leads to aggregation of EVs: (a) Western blot showing exogenous binding of GAPDH to HEK293T EVs. Increasing concentrations of histidine-(His6) and Flag-tagged GAPDH (lane 3,4 and 5) were incubated with a fixed number of EVs (see details in methods). Endogenous GAPDH (Endo. GAPDH) present naturally on the EV surface is shown in the top blot along with the exogenous GAPDH (Exo. GAPDH). Alix and CD81 are EV protein markers used as a positive control. Calnexin (bottom blot) was used to demonstrate the purity of EV samples. In this blot, lane 2, 3 and 4 represent EVs, and lane 5 represents cell lysate. Representative blots (n > 3). (b) UV-absorbance spectrum of EVs after passing through gel-filtration column. Increase in the absorbance of EVs+GAPDH peak indicates binding of GAPDH protein. Purified EVs were used for incubation with either GAPDH (upper chromatogram) or BSA protein (lower chromatogram). The first peak (at ∼10 ml elution) represents EVs and the second peak (around 20 ml elution) represents unbound proteins. Representative graphs (n > 3). (c) NTA profile showing the size distribution of purified HEK293T EVs after incubation with either GAPDH or BSA proteins respectively. Binding of GAPDH to the EVs, shifts their size. Inset is the scatter plot representing size (mean) of EVs (red; EVs+GAPDH, black; EVs+BSA). Data shown as mean ± s.d, n=9, ***p

    Techniques Used: Binding Assay, Western Blot, Incubation, Positive Control, Filtration, Purification

    2) Product Images from "Heat shock-induced activation of stress MAP kinase is regulated by threonine- and tyrosine-specific phosphatases"

    Article Title: Heat shock-induced activation of stress MAP kinase is regulated by threonine- and tyrosine-specific phosphatases

    Journal: Genes & Development

    doi:

    Ptc1 and Ptc3 phosphatases regulate phosphorylation of Spc1 Thr-171. ( A ) Wild-type (KS1376) and Δptc1Δptc3 mutant (CA135) strains carrying the spc1:HA6H allele were grown at 30°C to mid-log phase and shifted to 48°C at time 0. Cells were harvested at the indicated time points and Spc1 was purified on Ni–NTA beads for immunoblotting with anti-phospho-p38, anti-phosphotyrosine, and anti-HA antibodies. After the initial Spc1 activation, dual-phosphorylated Spc1 decreased more slowly in Δptc1 Δptc3 cells than in wild-type cells, a result that was reproduced in repeated experiments. ( B ) Wild-type spc1:HA6H strain (KS1376) was transformed with the pREP vector and the pREP- ptc3 + plasmid to express ptc3 + under the regulation of the thiamine-repressible nmt1 promoter. The transformants were grown at 30°C for 18 hr in EMM2 medium without thiamine to induce expression from the nmt1 promoter before the temperature shift to 48°C at time 0. Cells harvested at the indicated time points were subjected to analyses as described in A .
    Figure Legend Snippet: Ptc1 and Ptc3 phosphatases regulate phosphorylation of Spc1 Thr-171. ( A ) Wild-type (KS1376) and Δptc1Δptc3 mutant (CA135) strains carrying the spc1:HA6H allele were grown at 30°C to mid-log phase and shifted to 48°C at time 0. Cells were harvested at the indicated time points and Spc1 was purified on Ni–NTA beads for immunoblotting with anti-phospho-p38, anti-phosphotyrosine, and anti-HA antibodies. After the initial Spc1 activation, dual-phosphorylated Spc1 decreased more slowly in Δptc1 Δptc3 cells than in wild-type cells, a result that was reproduced in repeated experiments. ( B ) Wild-type spc1:HA6H strain (KS1376) was transformed with the pREP vector and the pREP- ptc3 + plasmid to express ptc3 + under the regulation of the thiamine-repressible nmt1 promoter. The transformants were grown at 30°C for 18 hr in EMM2 medium without thiamine to induce expression from the nmt1 promoter before the temperature shift to 48°C at time 0. Cells harvested at the indicated time points were subjected to analyses as described in A .

    Techniques Used: Mutagenesis, Purification, Activation Assay, Transformation Assay, Plasmid Preparation, Expressing

    Related Articles

    Binding Assay:

    Article Title: The mutational landscape of normal human endometrial epithelium
    Article Snippet: .. Following a 5 minute binding reaction and magnetic bead separation, genomic DNA was washed twice with 75% ethanol. ..

    Purification:

    Article Title: GAPDH controls extracellular vesicle biogenesis and enhances therapeutic potential of EVs in silencing the Huntingtin gene in mice via siRNA delivery
    Article Snippet: .. At the N-terminus of GAPDH, six residues of histidine were inserted for purification by Ni-NTA chromatography. ..

    Article Title: Heat shock-induced activation of stress MAP kinase is regulated by threonine- and tyrosine-specific phosphatases
    Article Snippet: .. Spc1HA6H protein was purified on Ni–NTA–agarose beads under denaturing conditions and analyzed by immunoblotting with anti-HA (12CA5), anti-phosphotyrosine, and anti-phospho-p38 MAPK (New England Biolabs) antibodies ( ). ..

    Chromatography:

    Article Title: GAPDH controls extracellular vesicle biogenesis and enhances therapeutic potential of EVs in silencing the Huntingtin gene in mice via siRNA delivery
    Article Snippet: .. At the N-terminus of GAPDH, six residues of histidine were inserted for purification by Ni-NTA chromatography. ..

    Incubation:

    Article Title: Antibody RING-Mediated Destruction of Endogenous Proteins
    Article Snippet: .. To remove the His6-MBP tag, fusion proteins were incubated with TEV protease, followed by Ni-NTA affinity chromatography to remove any uncleaved His6-MBP tagged proteins, free His6-MBP tag and TEV protease (also His6-tagged). ..

    Affinity Chromatography:

    Article Title: Antibody RING-Mediated Destruction of Endogenous Proteins
    Article Snippet: .. To remove the His6-MBP tag, fusion proteins were incubated with TEV protease, followed by Ni-NTA affinity chromatography to remove any uncleaved His6-MBP tagged proteins, free His6-MBP tag and TEV protease (also His6-tagged). ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    New England Biolabs ni nta chromatography
    G58T-mediated loading of siRNA on EVs: SDS-PAGE of the G58T-based proteins purified by <t>Ni-NTA</t> chromatography. Gel is stained with Coomassie brilliant blue dye. (b) Gel shift assay of G58T EVs (upper) and G58TF EVs (lower), reflecting binding of siRNA to EVs. 20pmoles of siRNA were incubated with the given number of EVs. Bands trapped near the wells represent bound siRNA. siRNA alone was used as negative control. Based on the gel shift assay, around 550 siRNAs bind to each G58T EV and 714 siRNAs to each G58TF EV. Higher binding of siRNA to G58TF is due to the arginine-rich FHV peptide. (c) RNase A protection assay of G58TF EVs bound to 50 pmoles of siRNA. Bands in the agarose gel represent siRNA isolated from G58TF EVs after treatment with RNase A (0.2 mg/ml). G58TF EVs bound to siRNA were incubated with RNase A for 6 h at 37°C. (d) Representative confocal microscopy images of N2a cells after 4h of treatment with G58TF EVs carrying FAM-labelled siRNA (green). Cells were treated with lysotracker red dye (red dots) to label late-endosomes. Inset in the merged figure represents magnified image of N2a cells showing colocalization of siRNA with lysotracker dye (yellow spots). Images were captured using 60X objective lens of Olympus confocal microscope FV1000. (e) <t>GAPDH</t> gene silencing by G58T EVs at mRNA level in N2a cells. Different amounts of GAPDH siRNA bound to G58T EVs were added to cells. Level of mRNA was quantified after 48h of treatment by probe-based real time PCR. Data were analysed using linear regression analysis and ΔΔCt method. Level of mRNA in control was assigned as 1 and used to determine the relative level of GAPDH mRNA in the treated cells. Results are shown as mean ± s.d, n= 3, ***p
    Ni Nta Chromatography, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ni nta chromatography/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ni nta chromatography - by Bioz Stars, 2021-08
    95/100 stars
      Buy from Supplier

    Image Search Results


    G58T-mediated loading of siRNA on EVs: SDS-PAGE of the G58T-based proteins purified by Ni-NTA chromatography. Gel is stained with Coomassie brilliant blue dye. (b) Gel shift assay of G58T EVs (upper) and G58TF EVs (lower), reflecting binding of siRNA to EVs. 20pmoles of siRNA were incubated with the given number of EVs. Bands trapped near the wells represent bound siRNA. siRNA alone was used as negative control. Based on the gel shift assay, around 550 siRNAs bind to each G58T EV and 714 siRNAs to each G58TF EV. Higher binding of siRNA to G58TF is due to the arginine-rich FHV peptide. (c) RNase A protection assay of G58TF EVs bound to 50 pmoles of siRNA. Bands in the agarose gel represent siRNA isolated from G58TF EVs after treatment with RNase A (0.2 mg/ml). G58TF EVs bound to siRNA were incubated with RNase A for 6 h at 37°C. (d) Representative confocal microscopy images of N2a cells after 4h of treatment with G58TF EVs carrying FAM-labelled siRNA (green). Cells were treated with lysotracker red dye (red dots) to label late-endosomes. Inset in the merged figure represents magnified image of N2a cells showing colocalization of siRNA with lysotracker dye (yellow spots). Images were captured using 60X objective lens of Olympus confocal microscope FV1000. (e) GAPDH gene silencing by G58T EVs at mRNA level in N2a cells. Different amounts of GAPDH siRNA bound to G58T EVs were added to cells. Level of mRNA was quantified after 48h of treatment by probe-based real time PCR. Data were analysed using linear regression analysis and ΔΔCt method. Level of mRNA in control was assigned as 1 and used to determine the relative level of GAPDH mRNA in the treated cells. Results are shown as mean ± s.d, n= 3, ***p

    Journal: bioRxiv

    Article Title: GAPDH controls extracellular vesicle biogenesis and enhances therapeutic potential of EVs in silencing the Huntingtin gene in mice via siRNA delivery

    doi: 10.1101/2020.01.09.899880

    Figure Lengend Snippet: G58T-mediated loading of siRNA on EVs: SDS-PAGE of the G58T-based proteins purified by Ni-NTA chromatography. Gel is stained with Coomassie brilliant blue dye. (b) Gel shift assay of G58T EVs (upper) and G58TF EVs (lower), reflecting binding of siRNA to EVs. 20pmoles of siRNA were incubated with the given number of EVs. Bands trapped near the wells represent bound siRNA. siRNA alone was used as negative control. Based on the gel shift assay, around 550 siRNAs bind to each G58T EV and 714 siRNAs to each G58TF EV. Higher binding of siRNA to G58TF is due to the arginine-rich FHV peptide. (c) RNase A protection assay of G58TF EVs bound to 50 pmoles of siRNA. Bands in the agarose gel represent siRNA isolated from G58TF EVs after treatment with RNase A (0.2 mg/ml). G58TF EVs bound to siRNA were incubated with RNase A for 6 h at 37°C. (d) Representative confocal microscopy images of N2a cells after 4h of treatment with G58TF EVs carrying FAM-labelled siRNA (green). Cells were treated with lysotracker red dye (red dots) to label late-endosomes. Inset in the merged figure represents magnified image of N2a cells showing colocalization of siRNA with lysotracker dye (yellow spots). Images were captured using 60X objective lens of Olympus confocal microscope FV1000. (e) GAPDH gene silencing by G58T EVs at mRNA level in N2a cells. Different amounts of GAPDH siRNA bound to G58T EVs were added to cells. Level of mRNA was quantified after 48h of treatment by probe-based real time PCR. Data were analysed using linear regression analysis and ΔΔCt method. Level of mRNA in control was assigned as 1 and used to determine the relative level of GAPDH mRNA in the treated cells. Results are shown as mean ± s.d, n= 3, ***p

    Article Snippet: At the N-terminus of GAPDH, six residues of histidine were inserted for purification by Ni-NTA chromatography.

    Techniques: SDS Page, Purification, Chromatography, Staining, Electrophoretic Mobility Shift Assay, Binding Assay, Incubation, Negative Control, Agarose Gel Electrophoresis, Isolation, Confocal Microscopy, Microscopy, Real-time Polymerase Chain Reaction

    Surface binding of GAPDH leads to aggregation of EVs: (a) Western blot showing exogenous binding of GAPDH to HEK293T EVs. Increasing concentrations of histidine-(His6) and Flag-tagged GAPDH (lane 3,4 and 5) were incubated with a fixed number of EVs (see details in methods). Endogenous GAPDH (Endo. GAPDH) present naturally on the EV surface is shown in the top blot along with the exogenous GAPDH (Exo. GAPDH). Alix and CD81 are EV protein markers used as a positive control. Calnexin (bottom blot) was used to demonstrate the purity of EV samples. In this blot, lane 2, 3 and 4 represent EVs, and lane 5 represents cell lysate. Representative blots (n > 3). (b) UV-absorbance spectrum of EVs after passing through gel-filtration column. Increase in the absorbance of EVs+GAPDH peak indicates binding of GAPDH protein. Purified EVs were used for incubation with either GAPDH (upper chromatogram) or BSA protein (lower chromatogram). The first peak (at ∼10 ml elution) represents EVs and the second peak (around 20 ml elution) represents unbound proteins. Representative graphs (n > 3). (c) NTA profile showing the size distribution of purified HEK293T EVs after incubation with either GAPDH or BSA proteins respectively. Binding of GAPDH to the EVs, shifts their size. Inset is the scatter plot representing size (mean) of EVs (red; EVs+GAPDH, black; EVs+BSA). Data shown as mean ± s.d, n=9, ***p

    Journal: bioRxiv

    Article Title: GAPDH controls extracellular vesicle biogenesis and enhances therapeutic potential of EVs in silencing the Huntingtin gene in mice via siRNA delivery

    doi: 10.1101/2020.01.09.899880

    Figure Lengend Snippet: Surface binding of GAPDH leads to aggregation of EVs: (a) Western blot showing exogenous binding of GAPDH to HEK293T EVs. Increasing concentrations of histidine-(His6) and Flag-tagged GAPDH (lane 3,4 and 5) were incubated with a fixed number of EVs (see details in methods). Endogenous GAPDH (Endo. GAPDH) present naturally on the EV surface is shown in the top blot along with the exogenous GAPDH (Exo. GAPDH). Alix and CD81 are EV protein markers used as a positive control. Calnexin (bottom blot) was used to demonstrate the purity of EV samples. In this blot, lane 2, 3 and 4 represent EVs, and lane 5 represents cell lysate. Representative blots (n > 3). (b) UV-absorbance spectrum of EVs after passing through gel-filtration column. Increase in the absorbance of EVs+GAPDH peak indicates binding of GAPDH protein. Purified EVs were used for incubation with either GAPDH (upper chromatogram) or BSA protein (lower chromatogram). The first peak (at ∼10 ml elution) represents EVs and the second peak (around 20 ml elution) represents unbound proteins. Representative graphs (n > 3). (c) NTA profile showing the size distribution of purified HEK293T EVs after incubation with either GAPDH or BSA proteins respectively. Binding of GAPDH to the EVs, shifts their size. Inset is the scatter plot representing size (mean) of EVs (red; EVs+GAPDH, black; EVs+BSA). Data shown as mean ± s.d, n=9, ***p

    Article Snippet: At the N-terminus of GAPDH, six residues of histidine were inserted for purification by Ni-NTA chromatography.

    Techniques: Binding Assay, Western Blot, Incubation, Positive Control, Filtration, Purification

    Ptc1 and Ptc3 phosphatases regulate phosphorylation of Spc1 Thr-171. ( A ) Wild-type (KS1376) and Δptc1Δptc3 mutant (CA135) strains carrying the spc1:HA6H allele were grown at 30°C to mid-log phase and shifted to 48°C at time 0. Cells were harvested at the indicated time points and Spc1 was purified on Ni–NTA beads for immunoblotting with anti-phospho-p38, anti-phosphotyrosine, and anti-HA antibodies. After the initial Spc1 activation, dual-phosphorylated Spc1 decreased more slowly in Δptc1 Δptc3 cells than in wild-type cells, a result that was reproduced in repeated experiments. ( B ) Wild-type spc1:HA6H strain (KS1376) was transformed with the pREP vector and the pREP- ptc3 + plasmid to express ptc3 + under the regulation of the thiamine-repressible nmt1 promoter. The transformants were grown at 30°C for 18 hr in EMM2 medium without thiamine to induce expression from the nmt1 promoter before the temperature shift to 48°C at time 0. Cells harvested at the indicated time points were subjected to analyses as described in A .

    Journal: Genes & Development

    Article Title: Heat shock-induced activation of stress MAP kinase is regulated by threonine- and tyrosine-specific phosphatases

    doi:

    Figure Lengend Snippet: Ptc1 and Ptc3 phosphatases regulate phosphorylation of Spc1 Thr-171. ( A ) Wild-type (KS1376) and Δptc1Δptc3 mutant (CA135) strains carrying the spc1:HA6H allele were grown at 30°C to mid-log phase and shifted to 48°C at time 0. Cells were harvested at the indicated time points and Spc1 was purified on Ni–NTA beads for immunoblotting with anti-phospho-p38, anti-phosphotyrosine, and anti-HA antibodies. After the initial Spc1 activation, dual-phosphorylated Spc1 decreased more slowly in Δptc1 Δptc3 cells than in wild-type cells, a result that was reproduced in repeated experiments. ( B ) Wild-type spc1:HA6H strain (KS1376) was transformed with the pREP vector and the pREP- ptc3 + plasmid to express ptc3 + under the regulation of the thiamine-repressible nmt1 promoter. The transformants were grown at 30°C for 18 hr in EMM2 medium without thiamine to induce expression from the nmt1 promoter before the temperature shift to 48°C at time 0. Cells harvested at the indicated time points were subjected to analyses as described in A .

    Article Snippet: Spc1HA6H protein was purified on Ni–NTA–agarose beads under denaturing conditions and analyzed by immunoblotting with anti-HA (12CA5), anti-phosphotyrosine, and anti-phospho-p38 MAPK (New England Biolabs) antibodies ( ).

    Techniques: Mutagenesis, Purification, Activation Assay, Transformation Assay, Plasmid Preparation, Expressing