Pglvh1

Manufactured by GenePharma

Sourced in China

The PGLVH1 is a laboratory instrument designed for general liquid handling applications. It provides precise and automated pipetting capabilities for a variety of liquid volumes. The core function of the PGLVH1 is to transfer liquids accurately and efficiently in a laboratory setting.

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Lab products found in correlation

Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Chloroform methanol, by Merck Group (1 mentions) Pcdna3, by Thermo Fisher Scientific (1 mentions) Sh nc, by GenePharma (1 mentions) Lv nc, by GenePharma (1 mentions)

4 protocols using pglvh1

Molecular Mechanisms of Lipid Regulation

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Short hairpin RNA (shRNA) targeted ZNF143, lncRNA NEAT1, ROCK2, or scrambled oligonucleotides were inserted into pGLVH1 (GenePharm, China) vector (shZNF143, shNEAT1, shROCK2). The full length of lncRNA NEAT1 or ROCK2 coding sequence was amplified and cloned into a pcDNA3.1 (Invitrogen, Carlsbad, CA, USA) vector (NEAT1, ROCK2). HepG2 and Huh-7 cells were transfected with the above plasmids or their corresponding controls using Lipofectamine 3000 (Invitrogen). After 48 h, cells were harvested for subsequently experiments.
Biochemical analysisChloroform-methanol extraction method was used to measure triglyceride (TG) levels in cells and liver tissues and total cholesterol (TC) levels in liver tissues as previous described [24 (link),25 (link)]. Briefly, collected cells and tissues were homogenized and then the lipids were extracted by homogenizing with chloroform: methanol (2:1 v/v) (Sigma-Aldrich). After drying and resuspending to obtain a homogeneous sample, TG and TC levels were measured using a biochemical analyser.
The serum index of liver tissues (alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels) were detected by their respectively kits purchased from Jiancheng (Nanjing, China) (ALT: C009-2-1; AST: C010-2-1) according to the instructions of manufacturer.

Dong Y., Hu M., Tan K, & Dai R. (2023). ZNF143 inhibits hepatocyte mitophagy and promotes non-alcoholic fatty liver disease by targeting increased lncRNA NEAT1 expression to activate ROCK2 pathway. Epigenetics, 18(1), 2239592.

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Lentiviral Overexpression and Knockdown of lncSSBP1

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A lentiviral vector containing human lncSSBP1 was purchased from GenePharma (Suzhou, China) and used to overexpression lncSSBP1 (referred to as Lv-lncSSBP1). The negative control lentivirus (Lv-NC) was also purchased from GenePharma. An shRNA lentivirus vector containing the target sequence of lncSSBP1 (5′-GGCGACAAGCAACAACAATCA-3′) was also used to knock down lncSSBP1 expression. The sequence was cloned into pGLVH1 (GenePharma, SuZhou, China) to generate Sh-lncSSBP1. A negative control lentivirus containing a non-targeting shRNA sequence (5′-TTCTCCGAACGTGTCACGT-3′; referred to as sh-NC) was used as a control. All cloned sequences were verified by automated sequencing (GenePharma, SuZhou, China).
All lentiviral vectors, including Lv-lncSSBP1, Lv-NC, sh-lncSSBP1 and sh-NC, were transfected into 293FT cells for packaging. The virus particles were harvested 72 h after transfection of 293FT cells. For stable transfection, BEAS-2B cells were grown in six-well plates to 50% confluence, and 1 mL of viral supernatant was added with 1 μL polybrene. The interference efficiency of the template was verified by RT-PCR analysis.

Li Y., Chen H., Li S., Li Y., Liu G., Bai J., Luo H., Lan X, & He Z. (2020). LncSSBP1 Functions as a Negative Regulator of IL-6 Through Interaction With hnRNPK in Bronchial Epithelial Cells Infected With Talaromyces marneffei. Frontiers in Immunology, 10, 2977.

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Knockdown of ATM, SIRT6, and HDM2 in Cells

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Briefly, cells were transfected with small interfering RNAs (siRNAs) for 48 hr using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer’s instructions. The siRNAs targeting human ATM, SIRT6, and HDM2 were purchased (GenePharma, China) with sequences as follows, si-ATM#1: 5’-AAUGUCUUUGAGUAGUAUGUU-3’ (Zhou et al., 2003 (link)); Si-ATM#2: 5’-AAGCACCAGUCCAGUAUUGGC-3’ (Zhang et al., 2005 (link)); si-SIRT6#1: 5’-AAGAAUGUGCCAAGUGUAAGA-3’; si-SIRT6#2: 5’-CCGGCTCTGCACCGTGGCTAA-3’; si-HDM2#1: 5’-AACGCCACAAATCTGATAGTA-3’; si-HDM2#2: 5’-AATGCCTCAATTCACATAGAT-3’. A scrambled siRNA sequence was used as control. Lentiviral shRNA constructs were generated in a pGLVH1 backbone (GenePharma, China), and virus was produced in HEK293 cells. To deplete ATM in HSF cells and SIRT6 in HepG2 cells, lentiviral infection was performed in the presence of 5 μg/ml polybrene. Two days later, the infected HSF cells or HepG2 cells were selected with 2 μg/ml puromycin. To downregulate sir-2.4 expression, the NL2099 worms were exposed to incubation plates containing HT115 bacteria with sir-2.4 RNAi vector.

Qian M., Liu Z., Peng L., Tang X., Meng F., Ao Y., Zhou M., Wang M., Cao X., Qin B., Wang Z., Zhou Z., Wang G., Gao Z., Xu J, & Liu B. (2018). Boosting ATM activity alleviates aging and extends lifespan in a mouse model of progeria. eLife, 7, e34836.

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Establish circSNX27-specific shRNA Cell Line

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CircSNX27-specific short hairpin RNA (shRNAs) (shcircSXN27-1, shcircSXN27-2, and shcircSXN27-3) were established by placing circSNX27 sequence into a lentivirus vector (pGLVH1, GenePharm, China). Transfected into HEK293T cells, and stably expressed cells were screened by puromycin (2 μg/mL). circSNX27 overexpression plasmid and miR-638 mimics, as well as their negative controls, were supplied by Hanheng Biotech (Shanghai, China).

Xi Y., Gai Y., Zhang W., Wang M., Liu Q, & Bi Y. (2024). Circular RNA SNX27 Facilitates Gastric Cancer Progression By Sponging miR-638. The Turkish Journal of Gastroenterology, 35(4), 280-287.

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