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mammalian expression vectors prk5-megfp-hmgb1-wt  (Addgene inc)


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    Structured Review

    Addgene inc mammalian expression vectors prk5-megfp-hmgb1-wt
    Mammalian Expression Vectors Prk5 Megfp Hmgb1 Wt, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/prk5+vector/prk5+ha+gst+raga+q66l/pm40468084-568-0-13
    Average 90 stars, based on 1 article reviews
    mammalian expression vectors prk5-megfp-hmgb1-wt - by Bioz Stars, 2026-09
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Differential HspBP1 expression accounts for the greater vulnerability of neurons than astrocytes to misfolded proteins
    Article Snippet: Htt-23Q, -73Q, and-130Q were generated by subcloning N-terminal fragments of huntingtin (1–230 amino acids) containing 23Q, 73Q, or 130Q into pDendra2-N (Clontech) using SalI and ApaI cloning sites. .. Murine HspBP1 was cloned into a pRK5 vector using BamH1 and EcoR1 cloning sites. sgRNA backbone vector and Cas9 plasmids were purchased from Addgene. ..

    Article Title: AMBRA1 Negatively Regulates the Function of ALDH1B1, a Cancer Stem Cell Marker, by Controlling Its Ubiquitination
    Article Snippet: .. Wild-type and mutant ubiquitins cloned into the pRK5 vector (pRK5-HA-Ub WT (Addgene plasmid 17608), K0(17603), K6(22900), K11(22901), K27(22902), K29(22903), K33(17607), K48(17605), K63(17606)) were kindly provided by Drs. Ted Dawson and Sandra Weller. .. Opti-MEM (Thermo Fisher Scientific, Waltham, MA, USA) and Lipofectamine 2000 transfection reagent (Invitrogen, Grand Island, NY, USA) were used for transgene transfection.

    Article Title: AMPK attenuates SHH subgroup medulloblastoma growth and metastasis by inhibiting NF-κB activation.
    Article Snippet: D283-Med cells (ATCC Cat# HTB-185, RRID:CVCL_1155) were cultured in MEM medium supplemented with 10% FBS. .. Full-length human PRKAA1 cDNA was generated by PCR and cloned into the pRK5 vector (RRID:Addgene_32693). siRNAs specific for the human PRKAA1, GLI1 and CCL5 were purchased from GenePharma (Shanghai, China). ..

    Article Title: Autophagy regulates spermatid differentiation via degradation of PDLIM1.
    Article Snippet: The rat anti-LAMP2 antibody (ab13524, Abcam) was kindly provided by Dr. Li Yu (Tsinghua University). .. Plasmids , cell culture and transfection Genes encoding FLAG-tagged PDLIM1, STMN1 (stathmin 1), PPP4R2 (protein phosphatase 4, regulatory subunit 2), NUDC (nudC nuclear distribution protein), MAP6 (microtubule-associated protein 6), YBX1 (Y box protein 1), and CD2AP (CD2-associated protein) were cloned into the SalI and NotI site of the modified pRK5 vector (AddGene, 71868; deposited by David Sabatini ). .. The plasmids were transfected using Lipofectamine 2000 (Invitrogen, 11668019) according to the manufacturer's instructions.

    Plasmid Preparation:

    Article Title: Differential HspBP1 expression accounts for the greater vulnerability of neurons than astrocytes to misfolded proteins
    Article Snippet: Htt-23Q, -73Q, and-130Q were generated by subcloning N-terminal fragments of huntingtin (1–230 amino acids) containing 23Q, 73Q, or 130Q into pDendra2-N (Clontech) using SalI and ApaI cloning sites. .. Murine HspBP1 was cloned into a pRK5 vector using BamH1 and EcoR1 cloning sites. sgRNA backbone vector and Cas9 plasmids were purchased from Addgene. ..

    Article Title: AMBRA1 Negatively Regulates the Function of ALDH1B1, a Cancer Stem Cell Marker, by Controlling Its Ubiquitination
    Article Snippet: .. Wild-type and mutant ubiquitins cloned into the pRK5 vector (pRK5-HA-Ub WT (Addgene plasmid 17608), K0(17603), K6(22900), K11(22901), K27(22902), K29(22903), K33(17607), K48(17605), K63(17606)) were kindly provided by Drs. Ted Dawson and Sandra Weller. .. Opti-MEM (Thermo Fisher Scientific, Waltham, MA, USA) and Lipofectamine 2000 transfection reagent (Invitrogen, Grand Island, NY, USA) were used for transgene transfection.

    Article Title: miR-27a-3p negatively regulates osteogenic differentiation of MC3T3-E1 preosteoblasts by targeting osterix
    Article Snippet: Primary antibodies against Osx (1:4,000; cat. no. ab94744; Abcam) and β-actin (1:1,000; cat. no. sc-47778; Santa Cruz Biotechnology, Inc.) were used. .. Osx cDNA was inserted into a pRK5 vector (Addgene, Inc.) at the Eco RI and Hind III sites to generate wild-type pRK5-Flag-Osx plasmid. ..

    Article Title: AMPK attenuates SHH subgroup medulloblastoma growth and metastasis by inhibiting NF-κB activation.
    Article Snippet: D283-Med cells (ATCC Cat# HTB-185, RRID:CVCL_1155) were cultured in MEM medium supplemented with 10% FBS. .. Full-length human PRKAA1 cDNA was generated by PCR and cloned into the pRK5 vector (RRID:Addgene_32693). siRNAs specific for the human PRKAA1, GLI1 and CCL5 were purchased from GenePharma (Shanghai, China). ..

    Article Title: Calpain Determines the Propensity of Adult Hippocampal Neural Stem Cells to Autophagic Cell Death Following Insulin Withdrawal.
    Article Snippet: .. pRK5-calpain 1 and -calpain 2 plasmids were constructed by subcloning the human calpain 1 (OriGene, Rockville, MD, http://www.origene.com; SC116897) and calpain 2 (OriGene; SC119079) genes into the pRK5 vector obtained from Addgene (plasmid 17612). .. For calpain knockdown experiments, HCN cells were transfected with SMARTpool small interfering RNAs (siRNAs) against rat calpain 1 (Dharmacon, Lafayette, CO, http://www.dharmacon.gelifesciences.com; L-092181-020005), calpain 2 (Dharmacon; L-092084-02-0005), or control nontargeting siRNAs (Dharmacon; D-001810-01-20).

    Article Title: Autophagy regulates spermatid differentiation via degradation of PDLIM1.
    Article Snippet: The rat anti-LAMP2 antibody (ab13524, Abcam) was kindly provided by Dr. Li Yu (Tsinghua University). .. Plasmids , cell culture and transfection Genes encoding FLAG-tagged PDLIM1, STMN1 (stathmin 1), PPP4R2 (protein phosphatase 4, regulatory subunit 2), NUDC (nudC nuclear distribution protein), MAP6 (microtubule-associated protein 6), YBX1 (Y box protein 1), and CD2AP (CD2-associated protein) were cloned into the SalI and NotI site of the modified pRK5 vector (AddGene, 71868; deposited by David Sabatini ). .. The plasmids were transfected using Lipofectamine 2000 (Invitrogen, 11668019) according to the manufacturer's instructions.

    Cloning:

    Article Title: Differential HspBP1 expression accounts for the greater vulnerability of neurons than astrocytes to misfolded proteins
    Article Snippet: Htt-23Q, -73Q, and-130Q were generated by subcloning N-terminal fragments of huntingtin (1–230 amino acids) containing 23Q, 73Q, or 130Q into pDendra2-N (Clontech) using SalI and ApaI cloning sites. .. Murine HspBP1 was cloned into a pRK5 vector using BamH1 and EcoR1 cloning sites. sgRNA backbone vector and Cas9 plasmids were purchased from Addgene. ..

    Mutagenesis:

    Article Title: AMBRA1 Negatively Regulates the Function of ALDH1B1, a Cancer Stem Cell Marker, by Controlling Its Ubiquitination
    Article Snippet: .. Wild-type and mutant ubiquitins cloned into the pRK5 vector (pRK5-HA-Ub WT (Addgene plasmid 17608), K0(17603), K6(22900), K11(22901), K27(22902), K29(22903), K33(17607), K48(17605), K63(17606)) were kindly provided by Drs. Ted Dawson and Sandra Weller. .. Opti-MEM (Thermo Fisher Scientific, Waltham, MA, USA) and Lipofectamine 2000 transfection reagent (Invitrogen, Grand Island, NY, USA) were used for transgene transfection.

    Construct:

    Article Title: Frizzled-4 C-terminus Distal to KTXXXW Motif is Essential for Normal Dishevelled Recruitment and Norrin-stimulated Activation of Lef/Tcf-dependent Transcriptional Activation
    Article Snippet: .. The mouse FZD 4 construct containing the V5-tag between F37 and G38 in a prk5 vector (Addgene, Cambridge, MA) was used to generate mutants employed in this study. .. The QuikChange Site-Directed Mutagenesis Kit (Agilent, Santa Clara, CA) was employed with specific primers to mutate codons.

    Article Title: Calpain Determines the Propensity of Adult Hippocampal Neural Stem Cells to Autophagic Cell Death Following Insulin Withdrawal.
    Article Snippet: .. pRK5-calpain 1 and -calpain 2 plasmids were constructed by subcloning the human calpain 1 (OriGene, Rockville, MD, http://www.origene.com; SC116897) and calpain 2 (OriGene; SC119079) genes into the pRK5 vector obtained from Addgene (plasmid 17612). .. For calpain knockdown experiments, HCN cells were transfected with SMARTpool small interfering RNAs (siRNAs) against rat calpain 1 (Dharmacon, Lafayette, CO, http://www.dharmacon.gelifesciences.com; L-092181-020005), calpain 2 (Dharmacon; L-092084-02-0005), or control nontargeting siRNAs (Dharmacon; D-001810-01-20).

    Generated:

    Article Title: AMPK attenuates SHH subgroup medulloblastoma growth and metastasis by inhibiting NF-κB activation.
    Article Snippet: D283-Med cells (ATCC Cat# HTB-185, RRID:CVCL_1155) were cultured in MEM medium supplemented with 10% FBS. .. Full-length human PRKAA1 cDNA was generated by PCR and cloned into the pRK5 vector (RRID:Addgene_32693). siRNAs specific for the human PRKAA1, GLI1 and CCL5 were purchased from GenePharma (Shanghai, China). ..

    Polymerase Chain Reaction:

    Article Title: AMPK attenuates SHH subgroup medulloblastoma growth and metastasis by inhibiting NF-κB activation.
    Article Snippet: D283-Med cells (ATCC Cat# HTB-185, RRID:CVCL_1155) were cultured in MEM medium supplemented with 10% FBS. .. Full-length human PRKAA1 cDNA was generated by PCR and cloned into the pRK5 vector (RRID:Addgene_32693). siRNAs specific for the human PRKAA1, GLI1 and CCL5 were purchased from GenePharma (Shanghai, China). ..

    Subcloning:

    Article Title: Calpain Determines the Propensity of Adult Hippocampal Neural Stem Cells to Autophagic Cell Death Following Insulin Withdrawal.
    Article Snippet: .. pRK5-calpain 1 and -calpain 2 plasmids were constructed by subcloning the human calpain 1 (OriGene, Rockville, MD, http://www.origene.com; SC116897) and calpain 2 (OriGene; SC119079) genes into the pRK5 vector obtained from Addgene (plasmid 17612). .. For calpain knockdown experiments, HCN cells were transfected with SMARTpool small interfering RNAs (siRNAs) against rat calpain 1 (Dharmacon, Lafayette, CO, http://www.dharmacon.gelifesciences.com; L-092181-020005), calpain 2 (Dharmacon; L-092084-02-0005), or control nontargeting siRNAs (Dharmacon; D-001810-01-20).

    Cell Culture:

    Article Title: Autophagy regulates spermatid differentiation via degradation of PDLIM1.
    Article Snippet: The rat anti-LAMP2 antibody (ab13524, Abcam) was kindly provided by Dr. Li Yu (Tsinghua University). .. Plasmids , cell culture and transfection Genes encoding FLAG-tagged PDLIM1, STMN1 (stathmin 1), PPP4R2 (protein phosphatase 4, regulatory subunit 2), NUDC (nudC nuclear distribution protein), MAP6 (microtubule-associated protein 6), YBX1 (Y box protein 1), and CD2AP (CD2-associated protein) were cloned into the SalI and NotI site of the modified pRK5 vector (AddGene, 71868; deposited by David Sabatini ). .. The plasmids were transfected using Lipofectamine 2000 (Invitrogen, 11668019) according to the manufacturer's instructions.

    Transfection:

    Article Title: Autophagy regulates spermatid differentiation via degradation of PDLIM1.
    Article Snippet: The rat anti-LAMP2 antibody (ab13524, Abcam) was kindly provided by Dr. Li Yu (Tsinghua University). .. Plasmids , cell culture and transfection Genes encoding FLAG-tagged PDLIM1, STMN1 (stathmin 1), PPP4R2 (protein phosphatase 4, regulatory subunit 2), NUDC (nudC nuclear distribution protein), MAP6 (microtubule-associated protein 6), YBX1 (Y box protein 1), and CD2AP (CD2-associated protein) were cloned into the SalI and NotI site of the modified pRK5 vector (AddGene, 71868; deposited by David Sabatini ). .. The plasmids were transfected using Lipofectamine 2000 (Invitrogen, 11668019) according to the manufacturer's instructions.

    Modification:

    Article Title: Autophagy regulates spermatid differentiation via degradation of PDLIM1.
    Article Snippet: The rat anti-LAMP2 antibody (ab13524, Abcam) was kindly provided by Dr. Li Yu (Tsinghua University). .. Plasmids , cell culture and transfection Genes encoding FLAG-tagged PDLIM1, STMN1 (stathmin 1), PPP4R2 (protein phosphatase 4, regulatory subunit 2), NUDC (nudC nuclear distribution protein), MAP6 (microtubule-associated protein 6), YBX1 (Y box protein 1), and CD2AP (CD2-associated protein) were cloned into the SalI and NotI site of the modified pRK5 vector (AddGene, 71868; deposited by David Sabatini ). .. The plasmids were transfected using Lipofectamine 2000 (Invitrogen, 11668019) according to the manufacturer's instructions.



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    Journal: Cell Reports Medicine

    Article Title: KRAS G12D -driven pentose phosphate pathway remodeling imparts a targetable vulnerability synergizing with MRTX1133 for durable remissions in PDAC

    doi: 10.1016/j.xcrm.2025.101966

    Figure Lengend Snippet:

    Article Snippet: In addition, the cDNA sequences of E2F1 and RING1 gene were cloned into the pRK5-HA vector. pCMV3-His-Rb (#HG10137-NH) was purchased from Sino Biological Inc. (Beijing, China). pRK5-HA-Ubi, pRK5- Ring1 , pRK5- UBE2T , deletions-mutant p53 plasmids were constructed in our previous article.

    Techniques: Mutagenesis, Recombinant, Virus, CCK-8 Assay, Membrane, Concentration Assay, Modification, Transfection, Agarose Gel Electrophoresis, Plasmid Preparation, Extraction, Bicinchoninic Acid Protein Assay, Staining, RNA Extraction, H&E Stain, Viability Assay, Reporter Assay, Labeling, Western Blot, Sequencing, Control, Software