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ptat2 2 expression vector  (Addgene inc)


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    Structured Review

    Addgene inc ptat2 2 expression vector
    Ptat2 2 Expression Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/35621/pTAT2%2E2+(Plasmid+%2335621)/pmc05056240-68-22-25
    Average 93 stars, based on 1 article reviews
    ptat2 2 expression vector - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Purification:

    Article Title: Discovering Antioxidant Molecules in the Archaea Domain: Peroxiredoxin Bcp1 from Sulfolobus solfataricus Protects H9c2 Cardiomyoblasts from Oxidative Stress
    Article Snippet: The bcp1 coding sequence was isolated from the pET30Bcp1 vector [ ] after digestion with the Nde I and Xho I restriction endonucleases. .. The bcp1 fragment was purified by agarose gel electrophoresis using the QIAquick Gel Extraction Kit (Qiagen, Milano, Italy) and cloned into the pTAT2.2 expression vector (Addgene, Cambridge, USA). .. E. coli BL21-CodonPlus (DE3)RIL competent cells were transformed with the obtained vector, pTAT2.2-Bcp1, and selected on LB agar plates with kanamycin (10 μ g mL −1 ) and chloramphenicol (34 μ g mL −1 ).

    Agarose Gel Electrophoresis:

    Article Title: Discovering Antioxidant Molecules in the Archaea Domain: Peroxiredoxin Bcp1 from Sulfolobus solfataricus Protects H9c2 Cardiomyoblasts from Oxidative Stress
    Article Snippet: The bcp1 coding sequence was isolated from the pET30Bcp1 vector [ ] after digestion with the Nde I and Xho I restriction endonucleases. .. The bcp1 fragment was purified by agarose gel electrophoresis using the QIAquick Gel Extraction Kit (Qiagen, Milano, Italy) and cloned into the pTAT2.2 expression vector (Addgene, Cambridge, USA). .. E. coli BL21-CodonPlus (DE3)RIL competent cells were transformed with the obtained vector, pTAT2.2-Bcp1, and selected on LB agar plates with kanamycin (10 μ g mL −1 ) and chloramphenicol (34 μ g mL −1 ).

    Gel Extraction:

    Article Title: Discovering Antioxidant Molecules in the Archaea Domain: Peroxiredoxin Bcp1 from Sulfolobus solfataricus Protects H9c2 Cardiomyoblasts from Oxidative Stress
    Article Snippet: The bcp1 coding sequence was isolated from the pET30Bcp1 vector [ ] after digestion with the Nde I and Xho I restriction endonucleases. .. The bcp1 fragment was purified by agarose gel electrophoresis using the QIAquick Gel Extraction Kit (Qiagen, Milano, Italy) and cloned into the pTAT2.2 expression vector (Addgene, Cambridge, USA). .. E. coli BL21-CodonPlus (DE3)RIL competent cells were transformed with the obtained vector, pTAT2.2-Bcp1, and selected on LB agar plates with kanamycin (10 μ g mL −1 ) and chloramphenicol (34 μ g mL −1 ).

    Clone Assay:

    Article Title: Discovering Antioxidant Molecules in the Archaea Domain: Peroxiredoxin Bcp1 from Sulfolobus solfataricus Protects H9c2 Cardiomyoblasts from Oxidative Stress
    Article Snippet: The bcp1 coding sequence was isolated from the pET30Bcp1 vector [ ] after digestion with the Nde I and Xho I restriction endonucleases. .. The bcp1 fragment was purified by agarose gel electrophoresis using the QIAquick Gel Extraction Kit (Qiagen, Milano, Italy) and cloned into the pTAT2.2 expression vector (Addgene, Cambridge, USA). .. E. coli BL21-CodonPlus (DE3)RIL competent cells were transformed with the obtained vector, pTAT2.2-Bcp1, and selected on LB agar plates with kanamycin (10 μ g mL −1 ) and chloramphenicol (34 μ g mL −1 ).

    Expressing:

    Article Title: Discovering Antioxidant Molecules in the Archaea Domain: Peroxiredoxin Bcp1 from Sulfolobus solfataricus Protects H9c2 Cardiomyoblasts from Oxidative Stress
    Article Snippet: The bcp1 coding sequence was isolated from the pET30Bcp1 vector [ ] after digestion with the Nde I and Xho I restriction endonucleases. .. The bcp1 fragment was purified by agarose gel electrophoresis using the QIAquick Gel Extraction Kit (Qiagen, Milano, Italy) and cloned into the pTAT2.2 expression vector (Addgene, Cambridge, USA). .. E. coli BL21-CodonPlus (DE3)RIL competent cells were transformed with the obtained vector, pTAT2.2-Bcp1, and selected on LB agar plates with kanamycin (10 μ g mL −1 ) and chloramphenicol (34 μ g mL −1 ).



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    (A,B) Immunoblots of HK-1 and <t>HK-2</t> protein (top) and quantified band intensities (bottom) in mitochondrial (A) and cytosolic fractions (B) of T47D cells following 50 min incubation in pH 7.8 or 6.2. HK band intensities of mitochondrial and cytosolic fractions were normalized for loading using COX4 and β-actin bands, respectively. Bars are mean ± SD of band intensities (n = 3) relative to cells in pH 6.2.
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    Image Search Results


    (A,B) Immunoblots of HK-1 and HK-2 protein (top) and quantified band intensities (bottom) in mitochondrial (A) and cytosolic fractions (B) of T47D cells following 50 min incubation in pH 7.8 or 6.2. HK band intensities of mitochondrial and cytosolic fractions were normalized for loading using COX4 and β-actin bands, respectively. Bars are mean ± SD of band intensities (n = 3) relative to cells in pH 6.2.

    Journal: PLoS ONE

    Article Title: Mild Alkalization Acutely Triggers the Warburg Effect by Enhancing Hexokinase Activity via Voltage-Dependent Anion Channel Binding

    doi: 10.1371/journal.pone.0159529

    Figure Lengend Snippet: (A,B) Immunoblots of HK-1 and HK-2 protein (top) and quantified band intensities (bottom) in mitochondrial (A) and cytosolic fractions (B) of T47D cells following 50 min incubation in pH 7.8 or 6.2. HK band intensities of mitochondrial and cytosolic fractions were normalized for loading using COX4 and β-actin bands, respectively. Bars are mean ± SD of band intensities (n = 3) relative to cells in pH 6.2.

    Article Snippet: Control siRNA (sc-37007), HK-1 siRNA (sc-39044), HK-2 siRNA (sc-35621), and VDAC1 siRNA (sc-42355) were from Santa Cruz Biotechnology (Dallas, TX).

    Techniques: Western Blot, Incubation

    Relative FDG uptake of T47D cells after transfection with specific siRNA against HK-1, HK-2, or VDAC1. Bars are mean ± SD of six samples from two separate experiments. †, p <0.0005; ‡, p <0.0001, compared to cells transfected with respective siRNA and incubated in pH 6.2.

    Journal: PLoS ONE

    Article Title: Mild Alkalization Acutely Triggers the Warburg Effect by Enhancing Hexokinase Activity via Voltage-Dependent Anion Channel Binding

    doi: 10.1371/journal.pone.0159529

    Figure Lengend Snippet: Relative FDG uptake of T47D cells after transfection with specific siRNA against HK-1, HK-2, or VDAC1. Bars are mean ± SD of six samples from two separate experiments. †, p <0.0005; ‡, p <0.0001, compared to cells transfected with respective siRNA and incubated in pH 6.2.

    Article Snippet: Control siRNA (sc-37007), HK-1 siRNA (sc-39044), HK-2 siRNA (sc-35621), and VDAC1 siRNA (sc-42355) were from Santa Cruz Biotechnology (Dallas, TX).

    Techniques: Transfection, Incubation

    (A,B) Confocal microscopy images of T47D cells double stained for VDAC and HK-1 (A) or VDAC and HK-2 (B) following incubation at pH 6.2 or 7.8.

    Journal: PLoS ONE

    Article Title: Mild Alkalization Acutely Triggers the Warburg Effect by Enhancing Hexokinase Activity via Voltage-Dependent Anion Channel Binding

    doi: 10.1371/journal.pone.0159529

    Figure Lengend Snippet: (A,B) Confocal microscopy images of T47D cells double stained for VDAC and HK-1 (A) or VDAC and HK-2 (B) following incubation at pH 6.2 or 7.8.

    Article Snippet: Control siRNA (sc-37007), HK-1 siRNA (sc-39044), HK-2 siRNA (sc-35621), and VDAC1 siRNA (sc-42355) were from Santa Cruz Biotechnology (Dallas, TX).

    Techniques: Confocal Microscopy, Staining, Incubation

    (A) Immunoblots (top) and quantified band intensities (bottom) of free HK-1 and HK-2 protein dissociated from isolated mitochondria into the supernatant following exposure to pH 6.2 or 7.8. (B) Immunoblots and quantified band intensities of HK-1 and HK-2 protein remaining in the mitochondrial pellets following pH exposure as above. Band intensities of HK from the mitochondrial pellet were normalized by COX4 bands. In supernatants, negative COX-4 bands confirmed the absence of mitochondria in the samples. Bars are mean ± SD of band intensities (n = 2) relative to that at pH 6.2 (A) or pH 7.8 (B).

    Journal: PLoS ONE

    Article Title: Mild Alkalization Acutely Triggers the Warburg Effect by Enhancing Hexokinase Activity via Voltage-Dependent Anion Channel Binding

    doi: 10.1371/journal.pone.0159529

    Figure Lengend Snippet: (A) Immunoblots (top) and quantified band intensities (bottom) of free HK-1 and HK-2 protein dissociated from isolated mitochondria into the supernatant following exposure to pH 6.2 or 7.8. (B) Immunoblots and quantified band intensities of HK-1 and HK-2 protein remaining in the mitochondrial pellets following pH exposure as above. Band intensities of HK from the mitochondrial pellet were normalized by COX4 bands. In supernatants, negative COX-4 bands confirmed the absence of mitochondria in the samples. Bars are mean ± SD of band intensities (n = 2) relative to that at pH 6.2 (A) or pH 7.8 (B).

    Article Snippet: Control siRNA (sc-37007), HK-1 siRNA (sc-39044), HK-2 siRNA (sc-35621), and VDAC1 siRNA (sc-42355) were from Santa Cruz Biotechnology (Dallas, TX).

    Techniques: Western Blot, Isolation

    (A, B) Immunoblots (top) and quantified band intensities (bottom) of recombinant HK-1 (A) and HK-2 (B) co-immunoprecipitated with purified VDAC protein. Recombinant HK and purified VDAC protein were pre-incubated in pH 6.2 or 7.8 for 50 min. Bars are mean ± SD (n = 3) of band intensities normalized by VDAC bands relative to that at pH 6.2.

    Journal: PLoS ONE

    Article Title: Mild Alkalization Acutely Triggers the Warburg Effect by Enhancing Hexokinase Activity via Voltage-Dependent Anion Channel Binding

    doi: 10.1371/journal.pone.0159529

    Figure Lengend Snippet: (A, B) Immunoblots (top) and quantified band intensities (bottom) of recombinant HK-1 (A) and HK-2 (B) co-immunoprecipitated with purified VDAC protein. Recombinant HK and purified VDAC protein were pre-incubated in pH 6.2 or 7.8 for 50 min. Bars are mean ± SD (n = 3) of band intensities normalized by VDAC bands relative to that at pH 6.2.

    Article Snippet: Control siRNA (sc-37007), HK-1 siRNA (sc-39044), HK-2 siRNA (sc-35621), and VDAC1 siRNA (sc-42355) were from Santa Cruz Biotechnology (Dallas, TX).

    Techniques: Western Blot, Recombinant, Immunoprecipitation, Purification, Incubation