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v4 10x genomics pn 1000691 primescript rt master mix takara bio rr036a xf cell mito stress test kit agilent technologies  (TaKaRa)


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    TaKaRa v4 10x genomics pn 1000691 primescript rt master mix takara bio rr036a xf cell mito stress test kit agilent technologies
    V4 10x Genomics Pn 1000691 Primescript Rt Master Mix Takara Bio Rr036a Xf Cell Mito Stress Test Kit Agilent Technologies, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 27062 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/test+kit/PrimeScript+RT+Master+Mix/pm42887499-260-80-88
    Average 99 stars, based on 27062 article reviews
    v4 10x genomics pn 1000691 primescript rt master mix takara bio rr036a xf cell mito stress test kit agilent technologies - by Bioz Stars, 2026-10
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    Reverse Transcription:

    Article Title: CYBB regulates synovial fibroblast inflammatory activation and sEV-mediated synovial-cartilage communication in osteoarthritis.
    Article Snippet: Total RNA was extracted from the cells using Trizol reagent (Invitrogen, Carlsbad, CA, USA). .. Subsequently, the RNA was reverse transcribed into complementary DNA (cDNA) utilizing the 5X PrimeScript RT Master Mix (Takara, Otsu, Shiga, Japan). .. The RT-PCR was conducted with a reaction mixture comprising 2 μl of cDNA, 0.5 μl of primers, 2 μl of double-distilled water (ddH2O) and 5 μl of TB Green PREMIX Ex Taq II (Takara, Otsu, Shiga, Japan).

    Article Title: Integrative multi-omics analysis defines a macrophage-associated phospholipid metabolism signature linked to immune remodeling and prognosis in lung adenocarcinoma.
    Article Snippet: .. Equal amounts of RNA were reverse transcribed into cDNA using PrimeScript RT Master Mix (Takara). .. RT-qPCR was performed using SYBR Premix Ex Taq (Takara) on an iQ5 Real-Time PCR Detection System (Bio-Rad).

    Article Title: Multimodal Programming of Extracellular Vesicles via Nano-Primed SECCS-Based 3D Microenvironments for Coordinated Ferroptosis Suppression and Myogenic Regeneration in Diabetes.
    Article Snippet: Chronic diabetic glucolipotoxicity is associated with progressive skeletal muscle atrophy, bone loss, cellular senescence, and ferroptotic injury.. Because ferroptosis inhibition alonemay not restore compromisedmyogenic capacity, we developed a combined stem cell-conditioning strategy integrating fullerol priming with the stem cell screen-enrich-combine(-biomaterials) circulating system (SECCS).. This integrated three-dimensional culture platform uses porous β-TCP granules and intermittent medium circulation to generate BMSCs-derived extracellular vesicles (EVs).

    Article Title: NEK2 promotes trastuzumab resistance in gastric cancer through WTAP/YTHDF1-dependent m6A regulation of FN1.
    Article Snippet: 1 Department of Gastroenterology, Lishui Municipal Central Hospital, The Fifth Affiliated Hospital of Wenzhou Medical University, Lishui 323000, Zhejiang, China 2 Gastrointestinal Surgery, Lishui Municipal Central Hospital, The Fifth Affiliated Hospital of Wenzhou Medical University, Lishui, Zhejiang, China Abstract Background Trastuzumab resistance remains a major challenge in the clinical management of gastric cancer (GC), and effective strategies to overcome resistance are still limited.. NEK2 has been implicated in GC progression and therapeutic resistance in several tumor types; however, its role in trastuzumab resistance in GC and the underlying mechanisms remain unclear.. Methods and Results Trastuzumab-resistant NCI-N87/TR cells were established from parental NCI-N87 cells by stepwise exposure to increasing concentrations of trastuzumab.

    Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its' 3'-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma.
    Article Snippet: Then, the slides were immersed into a series of the neutral buffer (90 mM Tris–HCl, 90 mM boric acid, and 2 mM EDTA, pH 8.0) at 25 V for 30 min and stained with 2.5 μg/ml PI (Sigma-Aldrich) for 15 min. Representative images were captured using a fluorescence microscope (Nikon). .. RNA isolation, qRT-PCR and RNA stability assay Total RNA was isolated from pancreas and PDAC cells using Trizol reagent (Invitrogen), and reverse transcribed using PrimeScript RT MasterMix (TaKaRa, Dalian, China). .. qRT-PCR was conducted using SYBR Green PCR MasterMix (ABI, Foster City, CA, USA) with ABI 7500 Real-Time PCR System (ABI).

    Synthesized:

    Article Title: Genome-Wide Identification and Characterization of the UMAMIT Gene Family in Brassica napus : Evolution, Expression, and Potential Association with Oil Content.
    Article Snippet: .. First-strand cDNA was synthesized using PrimeScript RT Master Mix (TaKaRa Biotech, Dalian, China) according to the manufacturer’s instructions. .. qRT-PCR reactions were performed on a CFX96 Touch Real-Time PCR detection system using SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA).

    Article Title: Selection and Validation of Reference Genes for RT-qPCR Across Different Organs, Flower Bud Developmental Stages, and Hormone Treatments in Ilex verticillata .
    Article Snippet: RNA integrity was assessed by electrophoresis on a 1.2% agarose gel, and RNA concentration and purity were quantified using a Thermo Scientific (NanoDrop 1000 spectrophotometer, Wilmington, DE, USA). .. First-strand cDNA was subsequently synthesized using the PrimerScript RT Master Mix (Perfect Real Time) (RR036A, Takara, Dalian, China), following the manufacturer’s instructions. ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: piR-823 and piR-20326 Differentially Regulate Stemness Programs in Ovarian Cancer Stem-like Cells.
    Article Snippet: .. Diluted RNAs (50ng/ul) and random primers were used according to the “PrimeScript RT Master Mix Synthesis System for RT-PCR (Takara Bio USA, Inc., San Jose, CA, USA)” (Table S3). .. “SsoAdvanced Universal SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA)” master mix was used for mRNA expression analysis of genes.

    Isolation:

    Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its' 3'-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma.
    Article Snippet: Then, the slides were immersed into a series of the neutral buffer (90 mM Tris–HCl, 90 mM boric acid, and 2 mM EDTA, pH 8.0) at 25 V for 30 min and stained with 2.5 μg/ml PI (Sigma-Aldrich) for 15 min. Representative images were captured using a fluorescence microscope (Nikon). .. RNA isolation, qRT-PCR and RNA stability assay Total RNA was isolated from pancreas and PDAC cells using Trizol reagent (Invitrogen), and reverse transcribed using PrimeScript RT MasterMix (TaKaRa, Dalian, China). .. qRT-PCR was conducted using SYBR Green PCR MasterMix (ABI, Foster City, CA, USA) with ABI 7500 Real-Time PCR System (ABI).

    Quantitative RT-PCR:

    Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its' 3'-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma.
    Article Snippet: Then, the slides were immersed into a series of the neutral buffer (90 mM Tris–HCl, 90 mM boric acid, and 2 mM EDTA, pH 8.0) at 25 V for 30 min and stained with 2.5 μg/ml PI (Sigma-Aldrich) for 15 min. Representative images were captured using a fluorescence microscope (Nikon). .. RNA isolation, qRT-PCR and RNA stability assay Total RNA was isolated from pancreas and PDAC cells using Trizol reagent (Invitrogen), and reverse transcribed using PrimeScript RT MasterMix (TaKaRa, Dalian, China). .. qRT-PCR was conducted using SYBR Green PCR MasterMix (ABI, Foster City, CA, USA) with ABI 7500 Real-Time PCR System (ABI).

    Stability:

    Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its' 3'-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma.
    Article Snippet: Then, the slides were immersed into a series of the neutral buffer (90 mM Tris–HCl, 90 mM boric acid, and 2 mM EDTA, pH 8.0) at 25 V for 30 min and stained with 2.5 μg/ml PI (Sigma-Aldrich) for 15 min. Representative images were captured using a fluorescence microscope (Nikon). .. RNA isolation, qRT-PCR and RNA stability assay Total RNA was isolated from pancreas and PDAC cells using Trizol reagent (Invitrogen), and reverse transcribed using PrimeScript RT MasterMix (TaKaRa, Dalian, China). .. qRT-PCR was conducted using SYBR Green PCR MasterMix (ABI, Foster City, CA, USA) with ABI 7500 Real-Time PCR System (ABI).



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    Genetic and Pharmacological Inhibition of Nupr1 Attenuates Ang II Induced Mitochondrial Apoptosis in VSMCs. (A) Cell ability was evaluated by CCK8 assay ( n = 8 per group). (B) Lactate production of indicated groups ( n = 6 per group). (C) Representative images of TEM in VSMCs from the indicated groups. (D) Quantification of mitochondrial size in VSMCs from indicated groups ( n = 6 per group). (E) Quantification of circularity index in VSMC from indicated groups ( n = 6 per group). (F) Real‐time monitoring of oxygen consumption rate (OCR) in indicated groups detected by O2k. (G–I) Quantification of ATP‐coupled respiration, mitochondrial basal respiration, and maximal respiration from indicated groups ( n = 6 per group). (J) Representative images of calcein and Annexin V fluorescence results showing mPTP opening and apoptosis in indicated groups. Stronger green fluorescence represents a lower degree of mPTP opening, while diminished fluorescence indicates increased mPTP opening. Stronger red fluorescence represents a higher apoptosis rate. (K, L) Quantification of fluorescence intensity, including Annexin V and Calcein ( n = 6 per group). (M) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (N) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group). (O) Quantification of maximal respiration from indicated groups ( n = 6 per group). (P) Real‐time monitoring of OCR in indicated groups detected by O2k. (Q, R) Quantification of ATP‐coupled respiration and mitochondrial basal respiration from indicated groups ( n = 6 per group). (S) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (T) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group).

    Journal: Advanced Science

    Article Title: Targeting Nupr1‐Mediated p53 Mitochondrial Sequestration Alleviates Aortic Dissection by Restoring the Gadd45b‐Dependent Survival Program

    doi: 10.1002/advs.78143

    Figure Lengend Snippet: Genetic and Pharmacological Inhibition of Nupr1 Attenuates Ang II Induced Mitochondrial Apoptosis in VSMCs. (A) Cell ability was evaluated by CCK8 assay ( n = 8 per group). (B) Lactate production of indicated groups ( n = 6 per group). (C) Representative images of TEM in VSMCs from the indicated groups. (D) Quantification of mitochondrial size in VSMCs from indicated groups ( n = 6 per group). (E) Quantification of circularity index in VSMC from indicated groups ( n = 6 per group). (F) Real‐time monitoring of oxygen consumption rate (OCR) in indicated groups detected by O2k. (G–I) Quantification of ATP‐coupled respiration, mitochondrial basal respiration, and maximal respiration from indicated groups ( n = 6 per group). (J) Representative images of calcein and Annexin V fluorescence results showing mPTP opening and apoptosis in indicated groups. Stronger green fluorescence represents a lower degree of mPTP opening, while diminished fluorescence indicates increased mPTP opening. Stronger red fluorescence represents a higher apoptosis rate. (K, L) Quantification of fluorescence intensity, including Annexin V and Calcein ( n = 6 per group). (M) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (N) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group). (O) Quantification of maximal respiration from indicated groups ( n = 6 per group). (P) Real‐time monitoring of OCR in indicated groups detected by O2k. (Q, R) Quantification of ATP‐coupled respiration and mitochondrial basal respiration from indicated groups ( n = 6 per group). (S) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (T) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group).

    Article Snippet: According to mPTP test kit instructions (C2009S, Beyotime), 200 μL of Calcein AM staining solution and fluorescence quenching working solution were added to each well after the respective cell treatments.

    Techniques: Inhibition, CCK-8 Assay, Real-time Monitoring, Fluorescence, Flow Cytometry, Analysis, Staining

    Gadd45b is Indispensable for the Protective Effects of Nupr1 Deficiency Against Mitochondrial Apoptosis. (A) Enrichment of KEGG pathways. (B) GSEA analysis in VSMCs treated with Ang II stimulation and Nupr1 deficiency. (C) Chordal diagram of GO pathways related with cell fate and the contributing DEPs. (D) Stem plot of DEPs in the apoptosis pathway from the indicated groups. (E) RT‐qPCR analysis of mRNA levels of candidate genes in VSMCs. Data were presented as relative fold change to controls ( n = 6 per group). (F) Expressions of Gadd45b in VSMCs measured by Western blot and its quantification from indicated groups measured by Western blot ( n = 5 per group). (G, H) Representative images of ROS staining in indicated groups. Semi‐quantitative analysis of mean fluorescence intensity in indicated groups ( n = 6 per group). (I) Representative images of calcein and Annexin V fluorescence results showing mPTP opening in indicated groups. (J) Quantification of fluorescence intensity, including Annexin V and mPTP ( n = 3 per group). (K) Co‐immunoprecipitation and Western blot assays of Myc‐Nupr1 and Gadd45b. (L) Co‐immunoprecipitation assays of Myc‐Gadd45b and Nupr1.

    Journal: Advanced Science

    Article Title: Targeting Nupr1‐Mediated p53 Mitochondrial Sequestration Alleviates Aortic Dissection by Restoring the Gadd45b‐Dependent Survival Program

    doi: 10.1002/advs.78143

    Figure Lengend Snippet: Gadd45b is Indispensable for the Protective Effects of Nupr1 Deficiency Against Mitochondrial Apoptosis. (A) Enrichment of KEGG pathways. (B) GSEA analysis in VSMCs treated with Ang II stimulation and Nupr1 deficiency. (C) Chordal diagram of GO pathways related with cell fate and the contributing DEPs. (D) Stem plot of DEPs in the apoptosis pathway from the indicated groups. (E) RT‐qPCR analysis of mRNA levels of candidate genes in VSMCs. Data were presented as relative fold change to controls ( n = 6 per group). (F) Expressions of Gadd45b in VSMCs measured by Western blot and its quantification from indicated groups measured by Western blot ( n = 5 per group). (G, H) Representative images of ROS staining in indicated groups. Semi‐quantitative analysis of mean fluorescence intensity in indicated groups ( n = 6 per group). (I) Representative images of calcein and Annexin V fluorescence results showing mPTP opening in indicated groups. (J) Quantification of fluorescence intensity, including Annexin V and mPTP ( n = 3 per group). (K) Co‐immunoprecipitation and Western blot assays of Myc‐Nupr1 and Gadd45b. (L) Co‐immunoprecipitation assays of Myc‐Gadd45b and Nupr1.

    Article Snippet: According to mPTP test kit instructions (C2009S, Beyotime), 200 μL of Calcein AM staining solution and fluorescence quenching working solution were added to each well after the respective cell treatments.

    Techniques: Analysis, Quantitative RT-PCR, Western Blot, Staining, Fluorescence, Co-Immunoprecipitation Assay, Significance Assay