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> Chemicals & Drugs > Amino Acid > Phosphoglycerate Kinase

Phosphoglycerate Kinase

Phosphoglycerate Kinase: A key enzyme in glycolysis that catalyzes the reversible phosphorylation of 3-phosphoglycerate to 1,3-bisphosphoglycerate.
This reaction is an important step in energy production and can be optimized to enhance reproducibility in research.
Explore AI-powered tools from PubCompare.ai to streamline your Phosphoglycerate Kinase studies, identify the most effective protocols, and boost productivity - all while maintaining a focus on reproducibility.

Most cited protocols related to «Phosphoglycerate Kinase»

1
Mammalian Expression of Recombinant Enzymes
The coding sequence of FLPo and ΦC31o was commercially synthesized de novo (Geneart GmbH, Regensburg, Germany) based on the published FLPe and ΦC31 coding sequence [8] (link), [11] (link). The coding sequence of endogenous ΦC31 was PCR-amplified from phage lysate (Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) GmbH, Braunschweig, Germany). A C-terminal SV40 nuclear localization signal was also added to the endogenous ΦC31 coding sequence as previously described [11] (link). The FLPo, ΦC31o, and ΦC31 coding sequences were blunt cloned into mammalian expression vectors driven by the high expressing phosphoglycerate kinase 1 (PGK) promoter and containing the bovine growth hormone polyadenylation (bpA) sequence.
Raymond C.S, & Soriano P. (2007). High-Efficiency FLP and ΦC31 Site-Specific Recombination in Mammalian Cells. PLoS ONE, 2(1), e162.
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Publication 2007
Bacteriophages Cloning Vectors Exons growth hormone, bovine Mammals N-fluoresceinylphosphatidylethanolamine Nuclear Localization Signals Open Reading Frames Phosphoglycerate Kinase Polyadenylation Simian virus 40
2
Generation of human induced pluripotent stem cells
The cDNAs encoding hOct4, hSox2, hKlf4 and c-myc (purchased from Open Biosystems) were subcloned into self-inactivating lentiviral vectors driven by the human phosphoglycerate kinase (PGK) promoter. Lentiviral vector supernatants were produced by triple co-transfection of the plasmid DNA encoding the vector, pCMVΔR8.91 and pUCMD.G into 293T cells. Human fetal lung fibroblasts (MRC-5) purchased from ATCC (CCL-171) were seeded at 1.5×104 cells/cm2 in Eagle's Minimum Essential Medium supplemented with 10% fetal bovine serum (FBS). The following day the fibroblasts were transduced with equal amounts of supernatants of the four lentiviral vectors in the presence of 4 ug/ml polybrene for ∼16 hours. Six days after transduction, fibroblasts were harvested by trypsinization and plated at 2×104 cells per 60 mm dish on a feeder layer of mytomycin C-treated mouse embryonic fibroblasts (CF-1). The next day, the medium was switched to hESC medium. The iPS lines were confirmed positive for Tra-1–81, Tra-1–60, SSEA-4 and Nanog by immunofluorescence and flow cytometry. In both hiPSC clones all 4 vector-encoded transgenes were found to be silenced.
Chambers S.M., Fasano C.A., Papapetrou E.P., Tomishima M., Sadelain M, & Studer L. (2009). Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nature biotechnology, 27(3), 275-280.
Publication 2009
Cells Clone Cells Cloning Vectors DNA, Complementary Embryo Feeder Cell Layers Fetal Bovine Serum Fetus Fibroblasts Flow Cytometry Fluorescent Antibody Technique Gastrin-Secreting Cells Homo sapiens Human Embryonic Stem Cells Human Induced Pluripotent Stem Cells Hyperostosis, Diffuse Idiopathic Skeletal Lung Mice, Laboratory Mitomycin Oncogenes, myc Phosphoglycerate Kinase Plasmids Polybrene stage-specific embryonic antigen-4 Transfection Transgenes
3
Generation of human induced pluripotent stem cells
The cDNAs encoding hOct4, hSox2, hKlf4 and c-myc (purchased from Open Biosystems) were subcloned into self-inactivating lentiviral vectors driven by the human phosphoglycerate kinase (PGK) promoter. Lentiviral vector supernatants were produced by triple co-transfection of the plasmid DNA encoding the vector, pCMVΔR8.91 and pUCMD.G into 293T cells. Human fetal lung fibroblasts (MRC-5) purchased from ATCC (CCL-171) were seeded at 1.5×104 cells/cm2 in Eagle's Minimum Essential Medium supplemented with 10% fetal bovine serum (FBS). The following day the fibroblasts were transduced with equal amounts of supernatants of the four lentiviral vectors in the presence of 4 ug/ml polybrene for ∼16 hours. Six days after transduction, fibroblasts were harvested by trypsinization and plated at 2×104 cells per 60 mm dish on a feeder layer of mytomycin C-treated mouse embryonic fibroblasts (CF-1). The next day, the medium was switched to hESC medium. The iPS lines were confirmed positive for Tra-1–81, Tra-1–60, SSEA-4 and Nanog by immunofluorescence and flow cytometry. In both hiPSC clones all 4 vector-encoded transgenes were found to be silenced.
Chambers S.M., Fasano C.A., Papapetrou E.P., Tomishima M., Sadelain M, & Studer L. (2009). Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nature biotechnology, 27(3), 275-280.
Publication 2009
Cells Clone Cells Cloning Vectors DNA, Complementary Embryo Feeder Cell Layers Fetal Bovine Serum Fetus Fibroblasts Flow Cytometry Fluorescent Antibody Technique Gastrin-Secreting Cells Homo sapiens Human Embryonic Stem Cells Human Induced Pluripotent Stem Cells Hyperostosis, Diffuse Idiopathic Skeletal Lung Mice, Laboratory Mitomycin Oncogenes, myc Phosphoglycerate Kinase Plasmids Polybrene stage-specific embryonic antigen-4 Transfection Transgenes
4
Construction and Packaging of pCCL-Empty Lentivirus
The pCCL-Empty plasmid was constructed from a previously characterised vector, MA1, in which two transgenes can be efficiently transcribed from a bi-directional origin: with the minimal cytomegalovirus promoter for ΔNGFR (a truncated form of the nerve growth factor receptor), and a second transgene transcribed from the human phosphoglycerate kinase promoter [15 (link)]. The pCCL-Empty plasmid was kindly provided by Prof M Levings [16 (link)]. The lentivirus packaging plasmids, psPAX2, pRSV-Rev, pMDLg/pRRE and the pMD2. G envelope plasmid containing VSV-G were kindly provided by D. Trono [4 (link)]. For lentivirus production, plasmids were prepared using the Endofree Plasmid Maxi kit (Qiagen) according to manufacturer’s protocol.
Cribbs A.P., Kennedy A., Gregory B, & Brennan F.M. (2013). Simplified production and concentration of lentiviral vectors to achieve high transduction in primary human T cells. BMC Biotechnology, 13, 98.
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Publication 2013
Cloning Vectors Cytomegalovirus Homo sapiens Lentivirus NGFR protein, human Phosphoglycerate Kinase Plasmids Reproduction Transgenes
5
Meiotic protein expression analysis
Cells from 5- to 10-ml aliquots of meiotic cultures were harvested, and 1 ml of 20% trichloroacetic acid (TCA) was added. The supernatant was removed after centrifugation, and the pellet was resuspended in 100 μl of 20% TCA and stored al −80°C. Samples were thawed on ice, glass beads were added, and cells were broken using a FastPrep FP120 cell disrupter (BIO 101 ThermoSavant, Obiogene, Carlsbad, CA). The lysate was recovered by punching a hole on the bottom of the tube, and the glass beads were further washed with 200 μl of 5% TCA. Lysates were centrifuged at 1000 × g for 3 min, and the pellet was thoroughly resuspended in 100 μl of 2× Laemmli buffer and 50 μl of 2 M Tris base. After boiling for 5 min, 10–20 μl were loaded in the gels. Ndt80 and Cdc5 production was analyzed in 10% SDS–PAGE gels with a 37.5:1 ratio of acrylamide:bisacrylamide. To resolve the phosphorylated forms of Mek1, 10% gels (acrylamide:bisacrylamide 29:1) containing 37.5 μM Phos-tag reagent and 75 μM MnCl2 were used. After running, Phos-tag gels were washed with 1 mM EDTA before transfer. The anti-Ndt80 (kindly provided by K. Benjamin) and anti-Cdc5 (sc-6733; Santa Cruz Biotechnology, Santa Cruz, CA) antibodies were used at 1:5000 and 1:500 dilutions, respectively. To detect phosphorylation of Cdc28 at tyrosine 19, anti–phospho-Cdc2(Tyr15) (Cell Signaling Technology, Beverly, MA) was used at 1:1000 dilution. Total Cdc28 was detected with anti-PSTAIRE (sc-53, 1:500 dilution; Santa Cruz Biotechnology,). The anti-Mek1 antibody was previously described (Refolio et al., 2011 ). Anti-tubulin (TAT1; 1:5000 dilution) and anti–phosphoglycerate kinase (PGK) (A-6457, 1:5000 dilution; Molecular Probes, Invitrogen, Carlsbad, CA) were used as loading controls. The ECL or ECL+ reagents (GE Healthcare, Piscataway, NJ) were used for detection. The signal was captured with a ChemiDoc XRS system (Bio-Rad, Hercules, CA), using the Quantity One software.
Acosta I., Ontoso D, & San-Segundo P.A. (2011). The budding yeast polo-like kinase Cdc5 regulates the Ndt80 branch of the meiotic recombination checkpoint pathway. Molecular Biology of the Cell, 22(18), 3478-3490.
Publication 2011
1,3-bis(bis(pyridin-2-ylmethyl)amino)propan-2-ol Acrylamide Antibodies Antibodies, Anti-Idiotypic CDC28 Protein Kinase, S cerevisiae CDK1 protein, human Cells Centrifugation Edetic Acid Gels Laemmli buffer manganese chloride MAP2K1 protein, human Miotics Molecular Probes Phosphoglycerate Kinase Phosphorylation SDS-PAGE Technique, Dilution Trichloroacetic Acid Tromethamine Tubulin Tyrosine

Most recents protocols related to «Phosphoglycerate Kinase»

1
Quantifying Wound-Induced Gene Expression
Unwounded leaves were collected immediately before wounding treatment and stored at -80°C. For wounding treatment, leaves from the same plants were pinched with a pair of tweezers as described by [7 (link)]. After 0.5 h and 1 h, the wounded leaves were collected sequentially. Leaves (unwounded, and wounded for 0.5 h and 1 h, all from the same plants) were ground in liquid N2 to fine powders with mortars and pestles. RNA was extracted using Total RNA Mini kit (plant) (Geneaid/FroggaBio, USA) with DNase treatment. First-strand cDNA was made using SuperScript IV reverse transcriptase (Invitrogen, USA) and a mixture of oligo dT and random primers according to the manufacturer’s instruction.
Quantitative real-time PCR (qPCR) using SYBR Green (Life Technologies, USA) was carried out to measure the relative KED transcription levels in leaves of tomato, Arabidopsis, corn, spruce and waterlily. Reference genes for ΔCt normalization were tomato phosphoglycerate kinase [8 (link)], Arabidopsis thaliana TIP41-like protein [9 (link)], corn Elongation factor 1-α [10 (link)], white spruce Elongation factor 1-α [11 (link)] and waterlily actin [12 (link)]. At least three plants from each species were tested. The primer sequences are listed in S1 Table in S1 File.
Zhang X.H., Swait D., Jin X.L., Vichyavichien P., Nifakos N., Kaplan N., Raymond L, & Harlin J.M. (2023). Evolutionary analysis of KED-rich proteins in plants. PLOS ONE, 18(3), e0279772.
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Publication 2023
Actins Arabidopsis Arabidopsis thalianas Deoxyribonuclease I DNA, Complementary Elongation Factor 1alpha Genes Lycopersicon esculentum Maize Nymphaea oligo (dT) Oligonucleotide Primers Phosphoglycerate Kinase Picea Plants Powder Proteins Real-Time Polymerase Chain Reaction RNA-Directed DNA Polymerase SYBR Green I Transcription, Genetic
2
Omentin-1 mRNA Expression in Adipose Tissue

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Publication 2023
Anabolism DNA, Complementary Fluorescein Genes, Housekeeping Homo sapiens ITLN1 protein, human Kidney Diseases Oligonucleotide Primers Phosphoglycerate Kinase Polymerase Chain Reaction Real-Time Polymerase Chain Reaction Reverse Transcription RNA, Messenger Tissue, Adipose
3
Quantification of Glycolytic Enzymes by ELISA
Hexokinase, 6-phosphofructokinase (PFK), phosphoglycerate kinase (PGK), and pyruvate kinase (CDC19) enzyme-linked immunosorbent assay (ELISA) kits were purchased from Shanghai Preferred Biotechnology Co., Ltd. Yeast cells were pretreated with Rg1, homogenates were centrifuged at 1620× g for 10 min, and the supernatant was taken to determine the amounts of hexokinase, PFK, PGK and CDC19. According to the instructions of the ELISA kits, 50 μL of standard was added to the standard well, and the sample wells contained 10 μL of sample and 40 μL of buffer. HRP-labeled antibody (100 μL) was added to the standard and sample wells, and the mixtures were incubated at 37 °C for 60 min. Washing was repeated five times. Then, 100 μL of substrate was added to each well, and the samples were incubated at 37 °C for 15 min in the dark. Stop solution (50 μL) was added, and the samples were detected at 450 nm using a microplate reader (Tecan Group).
Wang S., Qiao J., Jiang C., Pan D., Yu S., Chen J., Liu S., Zhang P., Zhao D, & Liu M. (2023). Ginsenoside Rg1 Delays Chronological Aging in a Yeast Model via CDC19- and SDH2-Mediated Cellular Metabolism. Antioxidants, 12(2), 296.
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Publication 2023
6-Phosphofructokinase Buffers Cells Enzyme-Linked Immunosorbent Assay Hexokinase Immunoglobulins MCM2 protein, human Phosphoglycerate Kinase Pyruvate Kinase Yeast, Dried
4
Generation and Characterization of Prx4 and Srx Knockout Mice
All schemes of mouse breeding and experimental protocols were reviewed and approved by the University of Kentucky Institutional Animal Care and Use Committee (UK protocol 2016–2306). All procedures on mice were conducted following the Policy on Humane Care and Use of Laboratory Animals, and the Guidelines of the Animal Care and Laboratory Animal Welfare (NIH). Mice, regardless of strain or genetic background, were all housed in standard cages in temperature-controlled environments under a 12-h light/12-h dark cycle with ad libitum access to standard chow and water unless otherwise indicated. Original Prx4−/− mice in the C57BL/6 background were originally established by Iuchi et al. [31 (link)]. Prx4−/− mice in pure FVB/N background were further established by cross breeding the C57BL/6 mice with FVB/N mice for multiple generations (>10). To obtain Prx4−/− female mice, Prx4 genomic DNA was cloned. In the first intron of Prx4, the phosphoglycerate kinase-driven neomycin cassette flanked by loxP sequences was inserted followed by an extra loxP sequence and introduced upstream of the first exon. Prx4−/y male mice were generated by breeding Prx4flox/+ female mice with male mice who express Cre recombinase as described before [31 (link)]. Srx−/− C57BL/6 mice [32 (link)], were first bred with FVB/N mice and after multiple generations (≥10) of backcross breeding, the offspring Srx−/− FVB mice were used to breed with Prx4−/− mice to generate Prx4/Srx double knockout (DKO) mice. The genotype of KO and DKO mice was confirmed by PCR-based mouse genotyping as described previously.
Hao Y., Jiang H., Thapa P., Ding N., Alshahrani A., Fujii J., Toledano M.B, & Wei Q. (2023). Critical Role of the Sulfiredoxin-Peroxiredoxin IV Axis in Urethane-Induced Non-Small Cell Lung Cancer. Antioxidants, 12(2), 367.
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Publication 2023
Animals Animals, Laboratory Cre recombinase Environment, Controlled Exons Females Genetic Background Genome Genotype Institutional Animal Care and Use Committees Introns Males Mice, House Mice, Inbred C57BL Neomycin Phosphoglycerate Kinase PRDX4 protein, human Strains
5
Protective mAbs against C. albicans
We identified and isolated a panel of protective mouse monoclonal antibodies (mAbs) specific for C. albicans cell surface peptides and a glycan epitope, and each mAb is protective in both immunocompetent mice and neutropenic mice. Therefore, we selected the following four conserved 14-mer protective peptide epitopes and one glycan epitope as targets for human IgGs: Fba (YGKDVKDLFDYAQE), Met6 (PRIGGQRELKKITE), Hwp1, and phosphoglycerate kinase 1 (Pgk1) (VPLDGKTITNNQRI). Each synthetic peptide was produced commercially (GenScript) and used as the coating antigen on enzyme immunoassay (EIA) plates. The purity of each peptide was >98.5%. The glycan epitope β-1,2-mannotriose (gift from David Bundle) was conjugated to bovine serum albumin (BSA) as a trimannose-BSA conjugate for EIA plate coating.
Xin H., Rosario-Colon J.A, & Eberle K. (2023). Novel Intravenous Immunoglobulin Therapy for the Prevention and Treatment of Candida auris and Candida albicans Disseminated Candidiasis. mSphere, 8(1), e00584-22.
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Publication 2023
Antigens Cells Enzyme Immunoassay Epitopes Homo sapiens Immunocompetence mannotriose Mus Peptides Phosphoglycerate Kinase polypeptide C Polysaccharides Serum Albumin, Bovine

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More about "Phosphoglycerate Kinase"

Phosphoglycerate kinase (PGK) is a crucial enzyme in the glycolytic pathway, catalyzing the reversible phosphorylation of 3-phosphoglycerate to 1,3-bisphosphoglycerate.
This reaction is a key step in energy production and can be optimized to enhance the reproducibility of your research.
Leveraging AI-powered tools like PubCompare.ai can streamline your PGK studies and help you identify the most effective protocols.
These innovative solutions allow you to quickly locate the best techniques from literature, preprints, and patents, using AI-driven comparisons to pinpoint the most efficient approaches.
By optimizing your PGK research protocols with PubCompare.ai, you can boost productivity and maintain a strong focus on reproducibility.
This is particularly important when working with related techniques and tools, such as the IScript cDNA synthesis kit, RNeasy Mini Kit, TRIzol reagent, QuantiTect Reverse Transcription Kit, High-Capacity cDNA Reverse Transcription Kit, Direct-zol RNA MiniPrep kit, ABI Prism 7000 Sequence Detection System, Maxima SYBR Green qPCR Master Mix kit, and Lipofectamine 2000.
Explore the full potential of PGK research by incorporating the insights and capabilities of PubCompare.ai into your workflow.
Streamline your studies, enhance reproducibility, and boost productivity - all while unlocking new discoveries in the field of glycolysis and energy production.