pcr primer strategy  (Millipore)


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    Name:
    PCR Primer
    Description:
    This second edition is a much praised and widely used manual that has been entirely revised and updated Each technique is presented with extensive background information advice and troubleshooting All current applications of PCR are covered in protocols that have the hallmark reliability of the previous edition
    Catalog Number:
    z701270
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    Structured Review

    Millipore pcr primer strategy
    PCR Primer
    This second edition is a much praised and widely used manual that has been entirely revised and updated Each technique is presented with extensive background information advice and troubleshooting All current applications of PCR are covered in protocols that have the hallmark reliability of the previous edition
    https://www.bioz.com/result/pcr primer strategy/product/Millipore
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcr primer strategy - by Bioz Stars, 2020-07
    95/100 stars

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    Related Articles

    Countercurrent Chromatography:

    Article Title: Differential Expression and Function of Stamp Family Proteins in Adipocyte Differentiation
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: Stamp1, forward 5′-ATA GGA AGT GGG GAT TTT GC-3′, reverse 5′-AGA TGT CTC AGG TCC CAC AA-3′; Stamp2 , forward 5′- TCA CTT CCT TGC CAT CAG-3′ , reverse 5′- GCT CCA CCT TAG AAT CGA AG-3′; Stamp3 , forward 5′- CCG TCC ATT GCT AAT TCC CTC-3′ , reverse 5′- CGG CAG GTA GAA CTT GTA GTG-3′ ; aP2 : forward 5′-GTC ACC ATC CGG TCA GAG AG-3′, reverse 5′-TCG ACT TTC CAT CCC ACT TC-3′; Pparγ , forward 5′-GCC CTT TGG TGA CTT TAT GG-3′, reverse 5′-GGC GGT CTC CAC TGA GAA TG-3′; C/ebpα , forward 5′-GCG GCA AAG CCA AGA AGT C-3′, reverse 5′-GCG GTC ATT GTC ACT GGT CA-3′; 36B4 , forward 5′-AAG CGC GCG TCC TGG CAT TGT CT-3′, reverse 5′-CCG CAG GGG CAG CAG TGG T-3′. .. Statistics Statistical analyses were performed using the Student’s t-test.

    Article Title: STAMP2 is required for human adipose-derived stem cell differentiation and adipocyte-facilitated prostate cancer growth in vivo
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: aP2 , forward 5′-TAC TGG GCC AGG AAT TTG AC-3′, reverse 5′-TGG TTG ATT TTC CAT CCC AT-3′; STEAP1, forward 5′-TTT GGT GCA AAT GCA AAA gc-3′, reverse AGG GTC AAG CTA AGG CGA AG-3′; STAMP1 , forward 5′-GCT CTT GTT TTG CCC TCA AT-3′, reverse 5′-GGG GAG ACA TGA GGA ATT GTT-3′; STAMP2 , forward 5′-ATG ACA GCA AAG CCA AGC AA-3′, reverse 5′-GCA AAG CAT CCA GTG GTC AA-3′; STAMP3 , forward 5′-GAG CAC ACT GCA CAC GCT CA-3′, reverse 5′CTC CCT CTC CCA GCC CTC TCC-3′; β-actin, forward 5′-GGC TAC AGC TTC ACC ACC AC-3′, reverse 5′-GTC AGG CAG CTC GTA GCT CT-3′; GAPDH , forward 5′-GTC AGT GGT GGA CCT GAC CT-3′, reverse 5′-GTC AGT GGT GGA CCT GAC CT-3′; GLUT4 , forward 5′-CAG ATC GGC TCT GAC GAT G-3′, reverse 5′-ACT GAA GGG AGC CAA GCA C-3′; PPARg , forward 5′-TCA AGA CAA CCT GCT ACA AGC CCT, reverse 5′-AAG AAG GGA AAt GTT GGC AGT GGC-3′; ACC1 , forward 5′-GTT GCA CAA AAG GAT TTC AG-3′, reverse 5′-CGC ATT ACC ATG CTC CGC AC-3′; GPAT1 , forward 5′-GAT GGC TTG CAA GAC GCC TTT CTT-3′, reverse 5′-GCC ACT TCT GCA ATT GCC TCT TGT-3′; CPT1 , forward 5′-TGG AGT CCC CTT TCC TTA AG-3′, reverse 5′-CCG TCA TCA GCA ACC G-3′; ACOX1 , forward 5′-GCT AAG AAC TCC CCA CTG AA-3′, reverse 5′- GAC ACT TCA GAG CTT GGA C -3′; IFIT1 , forward 5′-TTG CCT GGA TGT ATT ACC AC-3′, reverse 5′-GCT TCT TGC AAA TGT TCT CC-3′; IFIT3 , forward 5′-GAA CAT GCT GAC CAA GCA GA-3′, reverse 5′-CAG TTG TGT CCA CCC TTC CT-3′; MX1 , forward 5′-AGG ACC ATC GGA ATC TTG AC-3′, reverse 5′-TCA GGT CCA ACA CGA GGT TC-3′; XPO5 , forward 5′-TGT TAA CCC GAG AAG TCA TGG-3′, reverse 5′-GGT CTG TAA GCT CTG CCA TAG-3′; .. Western analysis ASCs were washed with PBS and whole cell extracts were prepared by resuspending cell pellets in 100 μl lysis buffer (20 mM HEPES (pH 7.7) [Sigma-Aldrich], 300 mM NaCl [Sigma-Aldrich], 1.5 mM MgCl2 [Sigma-Aldrich], 0.2 mM EDTA [Sigma-Aldrich], 0.1% Triton X-100 [Sigma-Aldrich], 0.5 mM dithiothreitol [Sigma-Aldrich] and a cocktail of protease inhibitors [Roche]) and incubated at 4°C for 1 hour.

    Real-time Polymerase Chain Reaction:

    Article Title: Regulation of transferrin receptor-1 mRNA by the interplay between IRE-binding proteins and miR-7/miR-141 in the 3′-IRE stem–loops
    Article Snippet: .. For mRNA expression analysis, total RNA was isolated using TRI Reagent RT (Molecular Research Center, Inc.). cDNA was reverse transcribed from 300 ng total RNA using the iScript cDNA Synthesis Kit (Bio-Rad) according to the manufacturer's protocol. cDNA was used in quantitative PCR (qPCR) to determine the expression of target genes. qPCR reagents (iTaq Universal SYBR Green Supermix) and thermocycler (CFX96) were from Bio-Rad. qPCR primer sequences were obtained from PrimerBank (TfR1, EGFR1) or realtimeprimers.com (GAPDH, HPRT1), and primer oligos were obtained from Sigma-Aldrich: TfR1 forward—5′-ACCATTGTCATATACCCGGTTCA-3′ TfR1 reverse—5′-CAATAGCCCAAGTAGCCAATCAT-3′ EGFR1 forward—5′-CCCCACCACGTACCAGATG-3′ EGFR1 reverse—5′-TCGCACTTCTTACACTTGCGG-3′ GAPDH forward—5′-GAGTCAACGGATTTGGTCGT-3′ GAPDH reverse—5′-TTGATTTTGGAGGGATCTCG-3′ HPRT1 forward—5′-TGACACTGGCAAAACAATGCA-3′ HPRT1 reverse —5′-GGTCCTTTTCACCAGCAAGCT-3′. .. IRP1 forward—5′-GATATGGGCGCTTACCATTTTCG-3′ IRP1 reverse—5′-TGTCGTCGCTGACATTCCAA-3′ IRP2 forward—5′-TCGATGTATCTAAACTTGGCACC-3′ IRP2 reverse—5′-GCCATCACAATTTCGTACAGCAG-3′ TBP forward—5′-TATAATCCCAAGCGGTTTGC-3′ TBP reverse —5′-GCTGGAAAACCCAACTTCTG-3′ TfR1 (mouse) forward—5′-GTTTCTGCCAGCCCCTTATT AT-3′ TfR1 (mouse) reverse—5′-GCAAGGAAAGGATATGCAGCA-3′ For microRNA quantitation, miR-7-5p, and miR-141-3p expression were measured by performing miRNA-specific reverse transcription (miScript RT II Kit, Qiagen) and quantitative PCR using miScript Primer Assays (miR-7: #MS00032116; miR-141: #MS00003507; U6 loading control: #MS00033740, Qiagen) according to the manufacturer's protocol.

    Article Title: Gene expression analysis of porcine whole blood cells infected with foot-and-mouth disease virus using high-throughput sequencing technology
    Article Snippet: .. Specific primers for 10 randomly selected DEGs were designed by the Sigma qPCR primer design online program and listed in . β-actin was used as an internal reference. .. The qPCR reactions were carried out in an ABI PRISM 7500 sequence detection system.

    Article Title: USP15 targets ALK3/BMPR1A for deubiquitylation to enhance bone morphogenetic protein signalling
    Article Snippet: .. RNAi and quantitative PCR The siRNA and qRT-PCR primer sequences used in this study are as follows: All siRNAs were purchased from Sigma. ..

    Isolation:

    Article Title: Regulation of transferrin receptor-1 mRNA by the interplay between IRE-binding proteins and miR-7/miR-141 in the 3′-IRE stem–loops
    Article Snippet: .. For mRNA expression analysis, total RNA was isolated using TRI Reagent RT (Molecular Research Center, Inc.). cDNA was reverse transcribed from 300 ng total RNA using the iScript cDNA Synthesis Kit (Bio-Rad) according to the manufacturer's protocol. cDNA was used in quantitative PCR (qPCR) to determine the expression of target genes. qPCR reagents (iTaq Universal SYBR Green Supermix) and thermocycler (CFX96) were from Bio-Rad. qPCR primer sequences were obtained from PrimerBank (TfR1, EGFR1) or realtimeprimers.com (GAPDH, HPRT1), and primer oligos were obtained from Sigma-Aldrich: TfR1 forward—5′-ACCATTGTCATATACCCGGTTCA-3′ TfR1 reverse—5′-CAATAGCCCAAGTAGCCAATCAT-3′ EGFR1 forward—5′-CCCCACCACGTACCAGATG-3′ EGFR1 reverse—5′-TCGCACTTCTTACACTTGCGG-3′ GAPDH forward—5′-GAGTCAACGGATTTGGTCGT-3′ GAPDH reverse—5′-TTGATTTTGGAGGGATCTCG-3′ HPRT1 forward—5′-TGACACTGGCAAAACAATGCA-3′ HPRT1 reverse —5′-GGTCCTTTTCACCAGCAAGCT-3′. .. IRP1 forward—5′-GATATGGGCGCTTACCATTTTCG-3′ IRP1 reverse—5′-TGTCGTCGCTGACATTCCAA-3′ IRP2 forward—5′-TCGATGTATCTAAACTTGGCACC-3′ IRP2 reverse—5′-GCCATCACAATTTCGTACAGCAG-3′ TBP forward—5′-TATAATCCCAAGCGGTTTGC-3′ TBP reverse —5′-GCTGGAAAACCCAACTTCTG-3′ TfR1 (mouse) forward—5′-GTTTCTGCCAGCCCCTTATT AT-3′ TfR1 (mouse) reverse—5′-GCAAGGAAAGGATATGCAGCA-3′ For microRNA quantitation, miR-7-5p, and miR-141-3p expression were measured by performing miRNA-specific reverse transcription (miScript RT II Kit, Qiagen) and quantitative PCR using miScript Primer Assays (miR-7: #MS00032116; miR-141: #MS00003507; U6 loading control: #MS00033740, Qiagen) according to the manufacturer's protocol.

    Quantitative RT-PCR:

    Article Title: Differential Expression and Function of Stamp Family Proteins in Adipocyte Differentiation
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: Stamp1, forward 5′-ATA GGA AGT GGG GAT TTT GC-3′, reverse 5′-AGA TGT CTC AGG TCC CAC AA-3′; Stamp2 , forward 5′- TCA CTT CCT TGC CAT CAG-3′ , reverse 5′- GCT CCA CCT TAG AAT CGA AG-3′; Stamp3 , forward 5′- CCG TCC ATT GCT AAT TCC CTC-3′ , reverse 5′- CGG CAG GTA GAA CTT GTA GTG-3′ ; aP2 : forward 5′-GTC ACC ATC CGG TCA GAG AG-3′, reverse 5′-TCG ACT TTC CAT CCC ACT TC-3′; Pparγ , forward 5′-GCC CTT TGG TGA CTT TAT GG-3′, reverse 5′-GGC GGT CTC CAC TGA GAA TG-3′; C/ebpα , forward 5′-GCG GCA AAG CCA AGA AGT C-3′, reverse 5′-GCG GTC ATT GTC ACT GGT CA-3′; 36B4 , forward 5′-AAG CGC GCG TCC TGG CAT TGT CT-3′, reverse 5′-CCG CAG GGG CAG CAG TGG T-3′. .. Statistics Statistical analyses were performed using the Student’s t-test.

    Article Title: USP15 targets ALK3/BMPR1A for deubiquitylation to enhance bone morphogenetic protein signalling
    Article Snippet: .. RNAi and quantitative PCR The siRNA and qRT-PCR primer sequences used in this study are as follows: All siRNAs were purchased from Sigma. ..

    Article Title: STAMP2 is required for human adipose-derived stem cell differentiation and adipocyte-facilitated prostate cancer growth in vivo
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: aP2 , forward 5′-TAC TGG GCC AGG AAT TTG AC-3′, reverse 5′-TGG TTG ATT TTC CAT CCC AT-3′; STEAP1, forward 5′-TTT GGT GCA AAT GCA AAA gc-3′, reverse AGG GTC AAG CTA AGG CGA AG-3′; STAMP1 , forward 5′-GCT CTT GTT TTG CCC TCA AT-3′, reverse 5′-GGG GAG ACA TGA GGA ATT GTT-3′; STAMP2 , forward 5′-ATG ACA GCA AAG CCA AGC AA-3′, reverse 5′-GCA AAG CAT CCA GTG GTC AA-3′; STAMP3 , forward 5′-GAG CAC ACT GCA CAC GCT CA-3′, reverse 5′CTC CCT CTC CCA GCC CTC TCC-3′; β-actin, forward 5′-GGC TAC AGC TTC ACC ACC AC-3′, reverse 5′-GTC AGG CAG CTC GTA GCT CT-3′; GAPDH , forward 5′-GTC AGT GGT GGA CCT GAC CT-3′, reverse 5′-GTC AGT GGT GGA CCT GAC CT-3′; GLUT4 , forward 5′-CAG ATC GGC TCT GAC GAT G-3′, reverse 5′-ACT GAA GGG AGC CAA GCA C-3′; PPARg , forward 5′-TCA AGA CAA CCT GCT ACA AGC CCT, reverse 5′-AAG AAG GGA AAt GTT GGC AGT GGC-3′; ACC1 , forward 5′-GTT GCA CAA AAG GAT TTC AG-3′, reverse 5′-CGC ATT ACC ATG CTC CGC AC-3′; GPAT1 , forward 5′-GAT GGC TTG CAA GAC GCC TTT CTT-3′, reverse 5′-GCC ACT TCT GCA ATT GCC TCT TGT-3′; CPT1 , forward 5′-TGG AGT CCC CTT TCC TTA AG-3′, reverse 5′-CCG TCA TCA GCA ACC G-3′; ACOX1 , forward 5′-GCT AAG AAC TCC CCA CTG AA-3′, reverse 5′- GAC ACT TCA GAG CTT GGA C -3′; IFIT1 , forward 5′-TTG CCT GGA TGT ATT ACC AC-3′, reverse 5′-GCT TCT TGC AAA TGT TCT CC-3′; IFIT3 , forward 5′-GAA CAT GCT GAC CAA GCA GA-3′, reverse 5′-CAG TTG TGT CCA CCC TTC CT-3′; MX1 , forward 5′-AGG ACC ATC GGA ATC TTG AC-3′, reverse 5′-TCA GGT CCA ACA CGA GGT TC-3′; XPO5 , forward 5′-TGT TAA CCC GAG AAG TCA TGG-3′, reverse 5′-GGT CTG TAA GCT CTG CCA TAG-3′; .. Western analysis ASCs were washed with PBS and whole cell extracts were prepared by resuspending cell pellets in 100 μl lysis buffer (20 mM HEPES (pH 7.7) [Sigma-Aldrich], 300 mM NaCl [Sigma-Aldrich], 1.5 mM MgCl2 [Sigma-Aldrich], 0.2 mM EDTA [Sigma-Aldrich], 0.1% Triton X-100 [Sigma-Aldrich], 0.5 mM dithiothreitol [Sigma-Aldrich] and a cocktail of protease inhibitors [Roche]) and incubated at 4°C for 1 hour.

    SYBR Green Assay:

    Article Title: Regulation of transferrin receptor-1 mRNA by the interplay between IRE-binding proteins and miR-7/miR-141 in the 3′-IRE stem–loops
    Article Snippet: .. For mRNA expression analysis, total RNA was isolated using TRI Reagent RT (Molecular Research Center, Inc.). cDNA was reverse transcribed from 300 ng total RNA using the iScript cDNA Synthesis Kit (Bio-Rad) according to the manufacturer's protocol. cDNA was used in quantitative PCR (qPCR) to determine the expression of target genes. qPCR reagents (iTaq Universal SYBR Green Supermix) and thermocycler (CFX96) were from Bio-Rad. qPCR primer sequences were obtained from PrimerBank (TfR1, EGFR1) or realtimeprimers.com (GAPDH, HPRT1), and primer oligos were obtained from Sigma-Aldrich: TfR1 forward—5′-ACCATTGTCATATACCCGGTTCA-3′ TfR1 reverse—5′-CAATAGCCCAAGTAGCCAATCAT-3′ EGFR1 forward—5′-CCCCACCACGTACCAGATG-3′ EGFR1 reverse—5′-TCGCACTTCTTACACTTGCGG-3′ GAPDH forward—5′-GAGTCAACGGATTTGGTCGT-3′ GAPDH reverse—5′-TTGATTTTGGAGGGATCTCG-3′ HPRT1 forward—5′-TGACACTGGCAAAACAATGCA-3′ HPRT1 reverse —5′-GGTCCTTTTCACCAGCAAGCT-3′. .. IRP1 forward—5′-GATATGGGCGCTTACCATTTTCG-3′ IRP1 reverse—5′-TGTCGTCGCTGACATTCCAA-3′ IRP2 forward—5′-TCGATGTATCTAAACTTGGCACC-3′ IRP2 reverse—5′-GCCATCACAATTTCGTACAGCAG-3′ TBP forward—5′-TATAATCCCAAGCGGTTTGC-3′ TBP reverse —5′-GCTGGAAAACCCAACTTCTG-3′ TfR1 (mouse) forward—5′-GTTTCTGCCAGCCCCTTATT AT-3′ TfR1 (mouse) reverse—5′-GCAAGGAAAGGATATGCAGCA-3′ For microRNA quantitation, miR-7-5p, and miR-141-3p expression were measured by performing miRNA-specific reverse transcription (miScript RT II Kit, Qiagen) and quantitative PCR using miScript Primer Assays (miR-7: #MS00032116; miR-141: #MS00003507; U6 loading control: #MS00033740, Qiagen) according to the manufacturer's protocol.

    Polymerase Chain Reaction:

    Article Title: Differential Expression and Function of Stamp Family Proteins in Adipocyte Differentiation
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: Stamp1, forward 5′-ATA GGA AGT GGG GAT TTT GC-3′, reverse 5′-AGA TGT CTC AGG TCC CAC AA-3′; Stamp2 , forward 5′- TCA CTT CCT TGC CAT CAG-3′ , reverse 5′- GCT CCA CCT TAG AAT CGA AG-3′; Stamp3 , forward 5′- CCG TCC ATT GCT AAT TCC CTC-3′ , reverse 5′- CGG CAG GTA GAA CTT GTA GTG-3′ ; aP2 : forward 5′-GTC ACC ATC CGG TCA GAG AG-3′, reverse 5′-TCG ACT TTC CAT CCC ACT TC-3′; Pparγ , forward 5′-GCC CTT TGG TGA CTT TAT GG-3′, reverse 5′-GGC GGT CTC CAC TGA GAA TG-3′; C/ebpα , forward 5′-GCG GCA AAG CCA AGA AGT C-3′, reverse 5′-GCG GTC ATT GTC ACT GGT CA-3′; 36B4 , forward 5′-AAG CGC GCG TCC TGG CAT TGT CT-3′, reverse 5′-CCG CAG GGG CAG CAG TGG T-3′. .. Statistics Statistical analyses were performed using the Student’s t-test.

    Article Title: STAMP2 is required for human adipose-derived stem cell differentiation and adipocyte-facilitated prostate cancer growth in vivo
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: aP2 , forward 5′-TAC TGG GCC AGG AAT TTG AC-3′, reverse 5′-TGG TTG ATT TTC CAT CCC AT-3′; STEAP1, forward 5′-TTT GGT GCA AAT GCA AAA gc-3′, reverse AGG GTC AAG CTA AGG CGA AG-3′; STAMP1 , forward 5′-GCT CTT GTT TTG CCC TCA AT-3′, reverse 5′-GGG GAG ACA TGA GGA ATT GTT-3′; STAMP2 , forward 5′-ATG ACA GCA AAG CCA AGC AA-3′, reverse 5′-GCA AAG CAT CCA GTG GTC AA-3′; STAMP3 , forward 5′-GAG CAC ACT GCA CAC GCT CA-3′, reverse 5′CTC CCT CTC CCA GCC CTC TCC-3′; β-actin, forward 5′-GGC TAC AGC TTC ACC ACC AC-3′, reverse 5′-GTC AGG CAG CTC GTA GCT CT-3′; GAPDH , forward 5′-GTC AGT GGT GGA CCT GAC CT-3′, reverse 5′-GTC AGT GGT GGA CCT GAC CT-3′; GLUT4 , forward 5′-CAG ATC GGC TCT GAC GAT G-3′, reverse 5′-ACT GAA GGG AGC CAA GCA C-3′; PPARg , forward 5′-TCA AGA CAA CCT GCT ACA AGC CCT, reverse 5′-AAG AAG GGA AAt GTT GGC AGT GGC-3′; ACC1 , forward 5′-GTT GCA CAA AAG GAT TTC AG-3′, reverse 5′-CGC ATT ACC ATG CTC CGC AC-3′; GPAT1 , forward 5′-GAT GGC TTG CAA GAC GCC TTT CTT-3′, reverse 5′-GCC ACT TCT GCA ATT GCC TCT TGT-3′; CPT1 , forward 5′-TGG AGT CCC CTT TCC TTA AG-3′, reverse 5′-CCG TCA TCA GCA ACC G-3′; ACOX1 , forward 5′-GCT AAG AAC TCC CCA CTG AA-3′, reverse 5′- GAC ACT TCA GAG CTT GGA C -3′; IFIT1 , forward 5′-TTG CCT GGA TGT ATT ACC AC-3′, reverse 5′-GCT TCT TGC AAA TGT TCT CC-3′; IFIT3 , forward 5′-GAA CAT GCT GAC CAA GCA GA-3′, reverse 5′-CAG TTG TGT CCA CCC TTC CT-3′; MX1 , forward 5′-AGG ACC ATC GGA ATC TTG AC-3′, reverse 5′-TCA GGT CCA ACA CGA GGT TC-3′; XPO5 , forward 5′-TGT TAA CCC GAG AAG TCA TGG-3′, reverse 5′-GGT CTG TAA GCT CTG CCA TAG-3′; .. Western analysis ASCs were washed with PBS and whole cell extracts were prepared by resuspending cell pellets in 100 μl lysis buffer (20 mM HEPES (pH 7.7) [Sigma-Aldrich], 300 mM NaCl [Sigma-Aldrich], 1.5 mM MgCl2 [Sigma-Aldrich], 0.2 mM EDTA [Sigma-Aldrich], 0.1% Triton X-100 [Sigma-Aldrich], 0.5 mM dithiothreitol [Sigma-Aldrich] and a cocktail of protease inhibitors [Roche]) and incubated at 4°C for 1 hour.

    Article Title: Bacterial Microbiome Dynamics in Post Pull-Through Hirschsprung-Associated Enterocolitis (HAEC): An Experimental Study Employing the Endothelin Receptor B-Null Mouse Model
    Article Snippet: .. PCRs were performed in an MJ Research PTC-200 thermal cycler (Bio-Rad Inc., Hercules, CA, USA) as 100 µl reactions containing: 50 mM Tris (pH 8.3), 500 µg/ml bovine serum albumin (BSA), 2.5 mM MgCl2 , 250 µM of each deoxynucleotide triphosphate (dNTP), 400 nM of the forward PCR primer, 200 nM of each reverse PCR primer, 2.5 µl of DNA template, and 0.625 units JumpStart Taq DNA polymerase (Sigma-Aldrich, St. Louis, MO, USA). .. PCR primers 515F (GTGCCAGCMGCCGCGGTAA) and 806R (GGACTACHVGGGTWTCTAAT) were used to targeted the 16S rRNA gene containing portions of the hypervariable regions V4 and V5, with the reverse primers including a 12 bp barcode ( ).

    Article Title: Phylogenetic analysis of LSU and SSU rDNA group I introns of lichen photobionts associated with the genera Xanthoria and Xanthomendoza (Teloschistaceae, lichenized Ascomycetes)
    Article Snippet: .. The PCR reactions were performed in 50 μL reaction volumes containing a reaction mix of 0.2 mM of each of four dNTPs, 1.5 μM of each PCR primer, and 1.25 U of Taq DNA polymerase (Sigma Aldrich, Buchs, SG, Switzerland) and 5 μL of 10X PCR buffer (100 mM Tris-HCl, pH 8.3, 500 mM KCl, 15 mM MgCl2 and 0.01% gelatin). .. Reactions were performed in a PTC 200 DNA engine (MJ Research Inc., Watertown, MA, USA) with the following conditions: initial denaturation for 3 min at 95°C, followed by 30 cycles of (30 sec at 95°C, 30 sec at 60°C, 1 min at 72°C) and a final extension for 10 min at 72°C.

    Expressing:

    Article Title: Regulation of transferrin receptor-1 mRNA by the interplay between IRE-binding proteins and miR-7/miR-141 in the 3′-IRE stem–loops
    Article Snippet: .. For mRNA expression analysis, total RNA was isolated using TRI Reagent RT (Molecular Research Center, Inc.). cDNA was reverse transcribed from 300 ng total RNA using the iScript cDNA Synthesis Kit (Bio-Rad) according to the manufacturer's protocol. cDNA was used in quantitative PCR (qPCR) to determine the expression of target genes. qPCR reagents (iTaq Universal SYBR Green Supermix) and thermocycler (CFX96) were from Bio-Rad. qPCR primer sequences were obtained from PrimerBank (TfR1, EGFR1) or realtimeprimers.com (GAPDH, HPRT1), and primer oligos were obtained from Sigma-Aldrich: TfR1 forward—5′-ACCATTGTCATATACCCGGTTCA-3′ TfR1 reverse—5′-CAATAGCCCAAGTAGCCAATCAT-3′ EGFR1 forward—5′-CCCCACCACGTACCAGATG-3′ EGFR1 reverse—5′-TCGCACTTCTTACACTTGCGG-3′ GAPDH forward—5′-GAGTCAACGGATTTGGTCGT-3′ GAPDH reverse—5′-TTGATTTTGGAGGGATCTCG-3′ HPRT1 forward—5′-TGACACTGGCAAAACAATGCA-3′ HPRT1 reverse —5′-GGTCCTTTTCACCAGCAAGCT-3′. .. IRP1 forward—5′-GATATGGGCGCTTACCATTTTCG-3′ IRP1 reverse—5′-TGTCGTCGCTGACATTCCAA-3′ IRP2 forward—5′-TCGATGTATCTAAACTTGGCACC-3′ IRP2 reverse—5′-GCCATCACAATTTCGTACAGCAG-3′ TBP forward—5′-TATAATCCCAAGCGGTTTGC-3′ TBP reverse —5′-GCTGGAAAACCCAACTTCTG-3′ TfR1 (mouse) forward—5′-GTTTCTGCCAGCCCCTTATT AT-3′ TfR1 (mouse) reverse—5′-GCAAGGAAAGGATATGCAGCA-3′ For microRNA quantitation, miR-7-5p, and miR-141-3p expression were measured by performing miRNA-specific reverse transcription (miScript RT II Kit, Qiagen) and quantitative PCR using miScript Primer Assays (miR-7: #MS00032116; miR-141: #MS00003507; U6 loading control: #MS00033740, Qiagen) according to the manufacturer's protocol.

    CTG Assay:

    Article Title: STAMP2 is required for human adipose-derived stem cell differentiation and adipocyte-facilitated prostate cancer growth in vivo
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: aP2 , forward 5′-TAC TGG GCC AGG AAT TTG AC-3′, reverse 5′-TGG TTG ATT TTC CAT CCC AT-3′; STEAP1, forward 5′-TTT GGT GCA AAT GCA AAA gc-3′, reverse AGG GTC AAG CTA AGG CGA AG-3′; STAMP1 , forward 5′-GCT CTT GTT TTG CCC TCA AT-3′, reverse 5′-GGG GAG ACA TGA GGA ATT GTT-3′; STAMP2 , forward 5′-ATG ACA GCA AAG CCA AGC AA-3′, reverse 5′-GCA AAG CAT CCA GTG GTC AA-3′; STAMP3 , forward 5′-GAG CAC ACT GCA CAC GCT CA-3′, reverse 5′CTC CCT CTC CCA GCC CTC TCC-3′; β-actin, forward 5′-GGC TAC AGC TTC ACC ACC AC-3′, reverse 5′-GTC AGG CAG CTC GTA GCT CT-3′; GAPDH , forward 5′-GTC AGT GGT GGA CCT GAC CT-3′, reverse 5′-GTC AGT GGT GGA CCT GAC CT-3′; GLUT4 , forward 5′-CAG ATC GGC TCT GAC GAT G-3′, reverse 5′-ACT GAA GGG AGC CAA GCA C-3′; PPARg , forward 5′-TCA AGA CAA CCT GCT ACA AGC CCT, reverse 5′-AAG AAG GGA AAt GTT GGC AGT GGC-3′; ACC1 , forward 5′-GTT GCA CAA AAG GAT TTC AG-3′, reverse 5′-CGC ATT ACC ATG CTC CGC AC-3′; GPAT1 , forward 5′-GAT GGC TTG CAA GAC GCC TTT CTT-3′, reverse 5′-GCC ACT TCT GCA ATT GCC TCT TGT-3′; CPT1 , forward 5′-TGG AGT CCC CTT TCC TTA AG-3′, reverse 5′-CCG TCA TCA GCA ACC G-3′; ACOX1 , forward 5′-GCT AAG AAC TCC CCA CTG AA-3′, reverse 5′- GAC ACT TCA GAG CTT GGA C -3′; IFIT1 , forward 5′-TTG CCT GGA TGT ATT ACC AC-3′, reverse 5′-GCT TCT TGC AAA TGT TCT CC-3′; IFIT3 , forward 5′-GAA CAT GCT GAC CAA GCA GA-3′, reverse 5′-CAG TTG TGT CCA CCC TTC CT-3′; MX1 , forward 5′-AGG ACC ATC GGA ATC TTG AC-3′, reverse 5′-TCA GGT CCA ACA CGA GGT TC-3′; XPO5 , forward 5′-TGT TAA CCC GAG AAG TCA TGG-3′, reverse 5′-GGT CTG TAA GCT CTG CCA TAG-3′; .. Western analysis ASCs were washed with PBS and whole cell extracts were prepared by resuspending cell pellets in 100 μl lysis buffer (20 mM HEPES (pH 7.7) [Sigma-Aldrich], 300 mM NaCl [Sigma-Aldrich], 1.5 mM MgCl2 [Sigma-Aldrich], 0.2 mM EDTA [Sigma-Aldrich], 0.1% Triton X-100 [Sigma-Aldrich], 0.5 mM dithiothreitol [Sigma-Aldrich] and a cocktail of protease inhibitors [Roche]) and incubated at 4°C for 1 hour.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Differential Expression and Function of Stamp Family Proteins in Adipocyte Differentiation
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: Stamp1, forward 5′-ATA GGA AGT GGG GAT TTT GC-3′, reverse 5′-AGA TGT CTC AGG TCC CAC AA-3′; Stamp2 , forward 5′- TCA CTT CCT TGC CAT CAG-3′ , reverse 5′- GCT CCA CCT TAG AAT CGA AG-3′; Stamp3 , forward 5′- CCG TCC ATT GCT AAT TCC CTC-3′ , reverse 5′- CGG CAG GTA GAA CTT GTA GTG-3′ ; aP2 : forward 5′-GTC ACC ATC CGG TCA GAG AG-3′, reverse 5′-TCG ACT TTC CAT CCC ACT TC-3′; Pparγ , forward 5′-GCC CTT TGG TGA CTT TAT GG-3′, reverse 5′-GGC GGT CTC CAC TGA GAA TG-3′; C/ebpα , forward 5′-GCG GCA AAG CCA AGA AGT C-3′, reverse 5′-GCG GTC ATT GTC ACT GGT CA-3′; 36B4 , forward 5′-AAG CGC GCG TCC TGG CAT TGT CT-3′, reverse 5′-CCG CAG GGG CAG CAG TGG T-3′. .. Statistics Statistical analyses were performed using the Student’s t-test.

    Article Title: STAMP2 is required for human adipose-derived stem cell differentiation and adipocyte-facilitated prostate cancer growth in vivo
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: aP2 , forward 5′-TAC TGG GCC AGG AAT TTG AC-3′, reverse 5′-TGG TTG ATT TTC CAT CCC AT-3′; STEAP1, forward 5′-TTT GGT GCA AAT GCA AAA gc-3′, reverse AGG GTC AAG CTA AGG CGA AG-3′; STAMP1 , forward 5′-GCT CTT GTT TTG CCC TCA AT-3′, reverse 5′-GGG GAG ACA TGA GGA ATT GTT-3′; STAMP2 , forward 5′-ATG ACA GCA AAG CCA AGC AA-3′, reverse 5′-GCA AAG CAT CCA GTG GTC AA-3′; STAMP3 , forward 5′-GAG CAC ACT GCA CAC GCT CA-3′, reverse 5′CTC CCT CTC CCA GCC CTC TCC-3′; β-actin, forward 5′-GGC TAC AGC TTC ACC ACC AC-3′, reverse 5′-GTC AGG CAG CTC GTA GCT CT-3′; GAPDH , forward 5′-GTC AGT GGT GGA CCT GAC CT-3′, reverse 5′-GTC AGT GGT GGA CCT GAC CT-3′; GLUT4 , forward 5′-CAG ATC GGC TCT GAC GAT G-3′, reverse 5′-ACT GAA GGG AGC CAA GCA C-3′; PPARg , forward 5′-TCA AGA CAA CCT GCT ACA AGC CCT, reverse 5′-AAG AAG GGA AAt GTT GGC AGT GGC-3′; ACC1 , forward 5′-GTT GCA CAA AAG GAT TTC AG-3′, reverse 5′-CGC ATT ACC ATG CTC CGC AC-3′; GPAT1 , forward 5′-GAT GGC TTG CAA GAC GCC TTT CTT-3′, reverse 5′-GCC ACT TCT GCA ATT GCC TCT TGT-3′; CPT1 , forward 5′-TGG AGT CCC CTT TCC TTA AG-3′, reverse 5′-CCG TCA TCA GCA ACC G-3′; ACOX1 , forward 5′-GCT AAG AAC TCC CCA CTG AA-3′, reverse 5′- GAC ACT TCA GAG CTT GGA C -3′; IFIT1 , forward 5′-TTG CCT GGA TGT ATT ACC AC-3′, reverse 5′-GCT TCT TGC AAA TGT TCT CC-3′; IFIT3 , forward 5′-GAA CAT GCT GAC CAA GCA GA-3′, reverse 5′-CAG TTG TGT CCA CCC TTC CT-3′; MX1 , forward 5′-AGG ACC ATC GGA ATC TTG AC-3′, reverse 5′-TCA GGT CCA ACA CGA GGT TC-3′; XPO5 , forward 5′-TGT TAA CCC GAG AAG TCA TGG-3′, reverse 5′-GGT CTG TAA GCT CTG CCA TAG-3′; .. Western analysis ASCs were washed with PBS and whole cell extracts were prepared by resuspending cell pellets in 100 μl lysis buffer (20 mM HEPES (pH 7.7) [Sigma-Aldrich], 300 mM NaCl [Sigma-Aldrich], 1.5 mM MgCl2 [Sigma-Aldrich], 0.2 mM EDTA [Sigma-Aldrich], 0.1% Triton X-100 [Sigma-Aldrich], 0.5 mM dithiothreitol [Sigma-Aldrich] and a cocktail of protease inhibitors [Roche]) and incubated at 4°C for 1 hour.

    Cellular Antioxidant Activity Assay:

    Article Title: STAMP2 is required for human adipose-derived stem cell differentiation and adipocyte-facilitated prostate cancer growth in vivo
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: aP2 , forward 5′-TAC TGG GCC AGG AAT TTG AC-3′, reverse 5′-TGG TTG ATT TTC CAT CCC AT-3′; STEAP1, forward 5′-TTT GGT GCA AAT GCA AAA gc-3′, reverse AGG GTC AAG CTA AGG CGA AG-3′; STAMP1 , forward 5′-GCT CTT GTT TTG CCC TCA AT-3′, reverse 5′-GGG GAG ACA TGA GGA ATT GTT-3′; STAMP2 , forward 5′-ATG ACA GCA AAG CCA AGC AA-3′, reverse 5′-GCA AAG CAT CCA GTG GTC AA-3′; STAMP3 , forward 5′-GAG CAC ACT GCA CAC GCT CA-3′, reverse 5′CTC CCT CTC CCA GCC CTC TCC-3′; β-actin, forward 5′-GGC TAC AGC TTC ACC ACC AC-3′, reverse 5′-GTC AGG CAG CTC GTA GCT CT-3′; GAPDH , forward 5′-GTC AGT GGT GGA CCT GAC CT-3′, reverse 5′-GTC AGT GGT GGA CCT GAC CT-3′; GLUT4 , forward 5′-CAG ATC GGC TCT GAC GAT G-3′, reverse 5′-ACT GAA GGG AGC CAA GCA C-3′; PPARg , forward 5′-TCA AGA CAA CCT GCT ACA AGC CCT, reverse 5′-AAG AAG GGA AAt GTT GGC AGT GGC-3′; ACC1 , forward 5′-GTT GCA CAA AAG GAT TTC AG-3′, reverse 5′-CGC ATT ACC ATG CTC CGC AC-3′; GPAT1 , forward 5′-GAT GGC TTG CAA GAC GCC TTT CTT-3′, reverse 5′-GCC ACT TCT GCA ATT GCC TCT TGT-3′; CPT1 , forward 5′-TGG AGT CCC CTT TCC TTA AG-3′, reverse 5′-CCG TCA TCA GCA ACC G-3′; ACOX1 , forward 5′-GCT AAG AAC TCC CCA CTG AA-3′, reverse 5′- GAC ACT TCA GAG CTT GGA C -3′; IFIT1 , forward 5′-TTG CCT GGA TGT ATT ACC AC-3′, reverse 5′-GCT TCT TGC AAA TGT TCT CC-3′; IFIT3 , forward 5′-GAA CAT GCT GAC CAA GCA GA-3′, reverse 5′-CAG TTG TGT CCA CCC TTC CT-3′; MX1 , forward 5′-AGG ACC ATC GGA ATC TTG AC-3′, reverse 5′-TCA GGT CCA ACA CGA GGT TC-3′; XPO5 , forward 5′-TGT TAA CCC GAG AAG TCA TGG-3′, reverse 5′-GGT CTG TAA GCT CTG CCA TAG-3′; .. Western analysis ASCs were washed with PBS and whole cell extracts were prepared by resuspending cell pellets in 100 μl lysis buffer (20 mM HEPES (pH 7.7) [Sigma-Aldrich], 300 mM NaCl [Sigma-Aldrich], 1.5 mM MgCl2 [Sigma-Aldrich], 0.2 mM EDTA [Sigma-Aldrich], 0.1% Triton X-100 [Sigma-Aldrich], 0.5 mM dithiothreitol [Sigma-Aldrich] and a cocktail of protease inhibitors [Roche]) and incubated at 4°C for 1 hour.

    Activated Clotting Time Assay:

    Article Title: Differential Expression and Function of Stamp Family Proteins in Adipocyte Differentiation
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: Stamp1, forward 5′-ATA GGA AGT GGG GAT TTT GC-3′, reverse 5′-AGA TGT CTC AGG TCC CAC AA-3′; Stamp2 , forward 5′- TCA CTT CCT TGC CAT CAG-3′ , reverse 5′- GCT CCA CCT TAG AAT CGA AG-3′; Stamp3 , forward 5′- CCG TCC ATT GCT AAT TCC CTC-3′ , reverse 5′- CGG CAG GTA GAA CTT GTA GTG-3′ ; aP2 : forward 5′-GTC ACC ATC CGG TCA GAG AG-3′, reverse 5′-TCG ACT TTC CAT CCC ACT TC-3′; Pparγ , forward 5′-GCC CTT TGG TGA CTT TAT GG-3′, reverse 5′-GGC GGT CTC CAC TGA GAA TG-3′; C/ebpα , forward 5′-GCG GCA AAG CCA AGA AGT C-3′, reverse 5′-GCG GTC ATT GTC ACT GGT CA-3′; 36B4 , forward 5′-AAG CGC GCG TCC TGG CAT TGT CT-3′, reverse 5′-CCG CAG GGG CAG CAG TGG T-3′. .. Statistics Statistical analyses were performed using the Student’s t-test.

    Article Title: STAMP2 is required for human adipose-derived stem cell differentiation and adipocyte-facilitated prostate cancer growth in vivo
    Article Snippet: .. All PCR products were analyzed by melting curve analysis. qRT-PCR primer sequences (all from Sigma-Aldrich) used in this study are as follows: aP2 , forward 5′-TAC TGG GCC AGG AAT TTG AC-3′, reverse 5′-TGG TTG ATT TTC CAT CCC AT-3′; STEAP1, forward 5′-TTT GGT GCA AAT GCA AAA gc-3′, reverse AGG GTC AAG CTA AGG CGA AG-3′; STAMP1 , forward 5′-GCT CTT GTT TTG CCC TCA AT-3′, reverse 5′-GGG GAG ACA TGA GGA ATT GTT-3′; STAMP2 , forward 5′-ATG ACA GCA AAG CCA AGC AA-3′, reverse 5′-GCA AAG CAT CCA GTG GTC AA-3′; STAMP3 , forward 5′-GAG CAC ACT GCA CAC GCT CA-3′, reverse 5′CTC CCT CTC CCA GCC CTC TCC-3′; β-actin, forward 5′-GGC TAC AGC TTC ACC ACC AC-3′, reverse 5′-GTC AGG CAG CTC GTA GCT CT-3′; GAPDH , forward 5′-GTC AGT GGT GGA CCT GAC CT-3′, reverse 5′-GTC AGT GGT GGA CCT GAC CT-3′; GLUT4 , forward 5′-CAG ATC GGC TCT GAC GAT G-3′, reverse 5′-ACT GAA GGG AGC CAA GCA C-3′; PPARg , forward 5′-TCA AGA CAA CCT GCT ACA AGC CCT, reverse 5′-AAG AAG GGA AAt GTT GGC AGT GGC-3′; ACC1 , forward 5′-GTT GCA CAA AAG GAT TTC AG-3′, reverse 5′-CGC ATT ACC ATG CTC CGC AC-3′; GPAT1 , forward 5′-GAT GGC TTG CAA GAC GCC TTT CTT-3′, reverse 5′-GCC ACT TCT GCA ATT GCC TCT TGT-3′; CPT1 , forward 5′-TGG AGT CCC CTT TCC TTA AG-3′, reverse 5′-CCG TCA TCA GCA ACC G-3′; ACOX1 , forward 5′-GCT AAG AAC TCC CCA CTG AA-3′, reverse 5′- GAC ACT TCA GAG CTT GGA C -3′; IFIT1 , forward 5′-TTG CCT GGA TGT ATT ACC AC-3′, reverse 5′-GCT TCT TGC AAA TGT TCT CC-3′; IFIT3 , forward 5′-GAA CAT GCT GAC CAA GCA GA-3′, reverse 5′-CAG TTG TGT CCA CCC TTC CT-3′; MX1 , forward 5′-AGG ACC ATC GGA ATC TTG AC-3′, reverse 5′-TCA GGT CCA ACA CGA GGT TC-3′; XPO5 , forward 5′-TGT TAA CCC GAG AAG TCA TGG-3′, reverse 5′-GGT CTG TAA GCT CTG CCA TAG-3′; .. Western analysis ASCs were washed with PBS and whole cell extracts were prepared by resuspending cell pellets in 100 μl lysis buffer (20 mM HEPES (pH 7.7) [Sigma-Aldrich], 300 mM NaCl [Sigma-Aldrich], 1.5 mM MgCl2 [Sigma-Aldrich], 0.2 mM EDTA [Sigma-Aldrich], 0.1% Triton X-100 [Sigma-Aldrich], 0.5 mM dithiothreitol [Sigma-Aldrich] and a cocktail of protease inhibitors [Roche]) and incubated at 4°C for 1 hour.

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  • 99
    Millipore 2 deoxyglucose
    Ube2o –/– mice show improved insulin sensitivity in diet-induced type 2 diabetes mouse model. ( A ) Blood glucose was measured Ube2o +/+ and Ube2o –/– mice on an HFD for 21 weeks. n = 6. ( B and C ) Glucose (GTT) ( B ) and insulin (ITT) ( C ) tolerance tests in Ube2o +/+ and Ube2o –/– mice on HFD for 21~22 weeks. Insets indicate AUC ( B ) and area above the curve (AAC) ( C ). Insulinemia at 15 minutes after the injection of glucose during a GTT test is shown in the right panel ( B ). Ube2o +/+ n = 6, Ube2o –/– n = 9. ( D – F ) Rates of basal and clamp endogenous glucose production ( D ), glucose infusion ( E ), and glucose disposal rate ( F ) during a hyperinsulinemic-euglycemic clamp study in Ube2o +/+ and Ube2o –/– mice. n = 6. ( G ) Insulin-stimulated 14 <t>C-2-deoxyglucose</t> uptake was assessed in visceral WAT and gastrocnemius muscle of Ube2o +/+ and Ube2o –/– mice during the final 45 minutes of the hyperinsulinemic-euglycemic clamps. n = 7. ( H ) Immunoblots (left) and statistical data (right) showing insulin-induced (200 nM) tyrosine phosphorylation of IRS1 immunoprecipitates and S473 phosphorylation of AKT protein and their total protein levels in the extensor digitorum longus muscle from Ube2o +/+ and Ube2o –/– mice on an HFD. n = 3. Error bars represent ± =SEM. P value was determined by Student’s t test (* P
    2 Deoxyglucose, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/2 deoxyglucose/product/Millipore
    Average 99 stars, based on 90 article reviews
    Price from $9.99 to $1999.99
    2 deoxyglucose - by Bioz Stars, 2020-07
    99/100 stars
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    89
    Millipore pcr strategy
    Ube2o –/– mice show improved insulin sensitivity in diet-induced type 2 diabetes mouse model. ( A ) Blood glucose was measured Ube2o +/+ and Ube2o –/– mice on an HFD for 21 weeks. n = 6. ( B and C ) Glucose (GTT) ( B ) and insulin (ITT) ( C ) tolerance tests in Ube2o +/+ and Ube2o –/– mice on HFD for 21~22 weeks. Insets indicate AUC ( B ) and area above the curve (AAC) ( C ). Insulinemia at 15 minutes after the injection of glucose during a GTT test is shown in the right panel ( B ). Ube2o +/+ n = 6, Ube2o –/– n = 9. ( D – F ) Rates of basal and clamp endogenous glucose production ( D ), glucose infusion ( E ), and glucose disposal rate ( F ) during a hyperinsulinemic-euglycemic clamp study in Ube2o +/+ and Ube2o –/– mice. n = 6. ( G ) Insulin-stimulated 14 <t>C-2-deoxyglucose</t> uptake was assessed in visceral WAT and gastrocnemius muscle of Ube2o +/+ and Ube2o –/– mice during the final 45 minutes of the hyperinsulinemic-euglycemic clamps. n = 7. ( H ) Immunoblots (left) and statistical data (right) showing insulin-induced (200 nM) tyrosine phosphorylation of IRS1 immunoprecipitates and S473 phosphorylation of AKT protein and their total protein levels in the extensor digitorum longus muscle from Ube2o +/+ and Ube2o –/– mice on an HFD. n = 3. Error bars represent ± =SEM. P value was determined by Student’s t test (* P
    Pcr Strategy, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr strategy/product/Millipore
    Average 89 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcr strategy - by Bioz Stars, 2020-07
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    Ube2o –/– mice show improved insulin sensitivity in diet-induced type 2 diabetes mouse model. ( A ) Blood glucose was measured Ube2o +/+ and Ube2o –/– mice on an HFD for 21 weeks. n = 6. ( B and C ) Glucose (GTT) ( B ) and insulin (ITT) ( C ) tolerance tests in Ube2o +/+ and Ube2o –/– mice on HFD for 21~22 weeks. Insets indicate AUC ( B ) and area above the curve (AAC) ( C ). Insulinemia at 15 minutes after the injection of glucose during a GTT test is shown in the right panel ( B ). Ube2o +/+ n = 6, Ube2o –/– n = 9. ( D – F ) Rates of basal and clamp endogenous glucose production ( D ), glucose infusion ( E ), and glucose disposal rate ( F ) during a hyperinsulinemic-euglycemic clamp study in Ube2o +/+ and Ube2o –/– mice. n = 6. ( G ) Insulin-stimulated 14 C-2-deoxyglucose uptake was assessed in visceral WAT and gastrocnemius muscle of Ube2o +/+ and Ube2o –/– mice during the final 45 minutes of the hyperinsulinemic-euglycemic clamps. n = 7. ( H ) Immunoblots (left) and statistical data (right) showing insulin-induced (200 nM) tyrosine phosphorylation of IRS1 immunoprecipitates and S473 phosphorylation of AKT protein and their total protein levels in the extensor digitorum longus muscle from Ube2o +/+ and Ube2o –/– mice on an HFD. n = 3. Error bars represent ± =SEM. P value was determined by Student’s t test (* P

    Journal: JCI Insight

    Article Title: A muscle-specific UBE2O/AMPKα2 axis promotes insulin resistance and metabolic syndrome in obesity

    doi: 10.1172/jci.insight.128269

    Figure Lengend Snippet: Ube2o –/– mice show improved insulin sensitivity in diet-induced type 2 diabetes mouse model. ( A ) Blood glucose was measured Ube2o +/+ and Ube2o –/– mice on an HFD for 21 weeks. n = 6. ( B and C ) Glucose (GTT) ( B ) and insulin (ITT) ( C ) tolerance tests in Ube2o +/+ and Ube2o –/– mice on HFD for 21~22 weeks. Insets indicate AUC ( B ) and area above the curve (AAC) ( C ). Insulinemia at 15 minutes after the injection of glucose during a GTT test is shown in the right panel ( B ). Ube2o +/+ n = 6, Ube2o –/– n = 9. ( D – F ) Rates of basal and clamp endogenous glucose production ( D ), glucose infusion ( E ), and glucose disposal rate ( F ) during a hyperinsulinemic-euglycemic clamp study in Ube2o +/+ and Ube2o –/– mice. n = 6. ( G ) Insulin-stimulated 14 C-2-deoxyglucose uptake was assessed in visceral WAT and gastrocnemius muscle of Ube2o +/+ and Ube2o –/– mice during the final 45 minutes of the hyperinsulinemic-euglycemic clamps. n = 7. ( H ) Immunoblots (left) and statistical data (right) showing insulin-induced (200 nM) tyrosine phosphorylation of IRS1 immunoprecipitates and S473 phosphorylation of AKT protein and their total protein levels in the extensor digitorum longus muscle from Ube2o +/+ and Ube2o –/– mice on an HFD. n = 3. Error bars represent ± =SEM. P value was determined by Student’s t test (* P

    Article Snippet: 2-Deoxyglucose, dexamethasone, etomoxir, FCCP, d -glucose, insulin, 3-isobutyl-1-methyl-xanthine, oligomycin, pioglitazone, and rotenone were purchased from MilliporeSigma.

    Techniques: Mouse Assay, Injection, Western Blot

    Regulation of AMPKα2 by skeletal muscle UBE2O. ( A ) UBE2O (top) or AMPKα2 (bottom) immunoprecipitates of lysates from skeletal muscle of control ( Ube2o fl/fl ) and Ube2o ΔMus mice were subjected to immunoblotting for AMPKα2 or UBE2O. ( B ) Lysates from C2C12 myotubes transfected with the indicated plasmids were subjected to metal affinity purification for His-tagged ubiquitin (His-Ub), then immunoblotting for ubiquitinated AMPKα2. Ni-NTA, Ni 2+ -nitrilotriacetic acid. ( C ) Recombinant AMPKα2 proteins were subjected to in vitro ubiquitination assay in the presence of in vitro translated WT or C1037S (CS) mutant UBE2O. ( D ) Lysates from C2C12 myotubes expressing WT or CS mutant UBE2O treated with MG132 (10 μM) for 6 hours were subjected to immunoblotting. ( E ) Lysates from C2C12 myotubes expressing WT or CS mutant UBE2O treated with cycloheximide (CHX) for the indicated times were subjected to immunoblotting (top). AMPKα2 or AMPKα1 protein levels were quantified by normalizing to the intensity of the HSP90 band (bottom). ( F ) Lysates from skeletal muscle of 12-week-old Ube2o +/+ , Ube2o –/– , Ube2o fl/fl , and Ube2o ΔMus mice were subjected to immunoblotting for the indicated proteins. ( G ) Lysates from C2C12 myotubes expressing UBE2O shRNA together with exogenous WT or CS mutant UBE2O from a Ube2o ORF transcript lacking the 3′ UTR sequence targeted by shRNA (shUbe2o-3′UTR) were subjected to immunoblotting for the indicated proteins. ( H ) Insulin-stimulated glucose uptake rate of C2C12 myotubes from G . 2-DG6P, 2-deoxyglucose-6-phosphate. n = 4. ( I ) Lysates from C2C12 myotubes expressing Ube2o shRNA together with Prkaa2 shRNA were subjected to immunoblotting for the indicated proteins. ( J ) Insulin-stimulated glucose uptake rate of C2C12 myotubes from I . n = 3. Error bars represent ±SEM. P value was determined by ANOVA (*** P

    Journal: JCI Insight

    Article Title: A muscle-specific UBE2O/AMPKα2 axis promotes insulin resistance and metabolic syndrome in obesity

    doi: 10.1172/jci.insight.128269

    Figure Lengend Snippet: Regulation of AMPKα2 by skeletal muscle UBE2O. ( A ) UBE2O (top) or AMPKα2 (bottom) immunoprecipitates of lysates from skeletal muscle of control ( Ube2o fl/fl ) and Ube2o ΔMus mice were subjected to immunoblotting for AMPKα2 or UBE2O. ( B ) Lysates from C2C12 myotubes transfected with the indicated plasmids were subjected to metal affinity purification for His-tagged ubiquitin (His-Ub), then immunoblotting for ubiquitinated AMPKα2. Ni-NTA, Ni 2+ -nitrilotriacetic acid. ( C ) Recombinant AMPKα2 proteins were subjected to in vitro ubiquitination assay in the presence of in vitro translated WT or C1037S (CS) mutant UBE2O. ( D ) Lysates from C2C12 myotubes expressing WT or CS mutant UBE2O treated with MG132 (10 μM) for 6 hours were subjected to immunoblotting. ( E ) Lysates from C2C12 myotubes expressing WT or CS mutant UBE2O treated with cycloheximide (CHX) for the indicated times were subjected to immunoblotting (top). AMPKα2 or AMPKα1 protein levels were quantified by normalizing to the intensity of the HSP90 band (bottom). ( F ) Lysates from skeletal muscle of 12-week-old Ube2o +/+ , Ube2o –/– , Ube2o fl/fl , and Ube2o ΔMus mice were subjected to immunoblotting for the indicated proteins. ( G ) Lysates from C2C12 myotubes expressing UBE2O shRNA together with exogenous WT or CS mutant UBE2O from a Ube2o ORF transcript lacking the 3′ UTR sequence targeted by shRNA (shUbe2o-3′UTR) were subjected to immunoblotting for the indicated proteins. ( H ) Insulin-stimulated glucose uptake rate of C2C12 myotubes from G . 2-DG6P, 2-deoxyglucose-6-phosphate. n = 4. ( I ) Lysates from C2C12 myotubes expressing Ube2o shRNA together with Prkaa2 shRNA were subjected to immunoblotting for the indicated proteins. ( J ) Insulin-stimulated glucose uptake rate of C2C12 myotubes from I . n = 3. Error bars represent ±SEM. P value was determined by ANOVA (*** P

    Article Snippet: 2-Deoxyglucose, dexamethasone, etomoxir, FCCP, d -glucose, insulin, 3-isobutyl-1-methyl-xanthine, oligomycin, pioglitazone, and rotenone were purchased from MilliporeSigma.

    Techniques: Mouse Assay, Transfection, Affinity Purification, Recombinant, In Vitro, Ubiquitin Assay, Mutagenesis, Expressing, shRNA, Sequencing

    UBE2O is upregulated in obese subjects with type 2 diabetes. ( A ) An RNA interference (RNAi) screen identifies UBE2O as a potent regulator of glucose uptake in myotubes. Primary normal human skeletal myotubes (HSkMs) and C2C12 mouse myotubes expressing a nontargeting control siRNA pool (Cont. siRNAs) or a synthetic siRNA library targeting 30 human and mouse E2s, respectively, were subjected to a screen for insulin-stimulated 2-deoxyglucose uptake. 2-DG6P, 2-deoxyglucose-6-phosphate. n = 3. ( B ) Total RNAs from quadriceps skeletal muscles of mice fed a normal chow or an HFD for 28 weeks were subjected to RT-qPCR. n = 4. ( C and D ) Lysates from quadriceps skeletal muscles of 9-month-old Ube2o +/+ mice fed normal chow or an HFD for 28 weeks ( C ) and 25-week-old type 2 diabetic db/db and lean ( db/+ ) mice ( D ) were subjected to immunoblotting. Error bars represent ±SEM. P value was determined by Student’s t test. * P

    Journal: JCI Insight

    Article Title: A muscle-specific UBE2O/AMPKα2 axis promotes insulin resistance and metabolic syndrome in obesity

    doi: 10.1172/jci.insight.128269

    Figure Lengend Snippet: UBE2O is upregulated in obese subjects with type 2 diabetes. ( A ) An RNA interference (RNAi) screen identifies UBE2O as a potent regulator of glucose uptake in myotubes. Primary normal human skeletal myotubes (HSkMs) and C2C12 mouse myotubes expressing a nontargeting control siRNA pool (Cont. siRNAs) or a synthetic siRNA library targeting 30 human and mouse E2s, respectively, were subjected to a screen for insulin-stimulated 2-deoxyglucose uptake. 2-DG6P, 2-deoxyglucose-6-phosphate. n = 3. ( B ) Total RNAs from quadriceps skeletal muscles of mice fed a normal chow or an HFD for 28 weeks were subjected to RT-qPCR. n = 4. ( C and D ) Lysates from quadriceps skeletal muscles of 9-month-old Ube2o +/+ mice fed normal chow or an HFD for 28 weeks ( C ) and 25-week-old type 2 diabetic db/db and lean ( db/+ ) mice ( D ) were subjected to immunoblotting. Error bars represent ±SEM. P value was determined by Student’s t test. * P

    Article Snippet: 2-Deoxyglucose, dexamethasone, etomoxir, FCCP, d -glucose, insulin, 3-isobutyl-1-methyl-xanthine, oligomycin, pioglitazone, and rotenone were purchased from MilliporeSigma.

    Techniques: Expressing, Mouse Assay, Quantitative RT-PCR