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Cell Biology

protocols
Protocol

Strategy For Constructing Gene Knock-in Cell Lines

Experiment Tags: Knock-in Cell Lines, Knock-in

Experiment Summary: Because the success of knock-in cell line construction is influenced by various factors, including the length of the knock-in gene fragment, homologous recombination efficiency, and cell line type, which can impact editing efficiency, this also makes the knock-in technology challenging. This article introduces the principles of constructing KI cell lines and the details that need to be paid attention to.

protocols
Protocol

Mammalian cell subculture

Experiment Tags: Mammalian cell, cell subculture

Experiment Summary: The basic principles, experimental steps and precautions for subculture of HEK293 cells and CHO cells will be introduced.

protocols
Protocol

Comparison of Cell Transfection Methods

Experiment Tags: Cell Transfection, Liposome Transfection

Experiment Summary: For mammalian cell transfection and protein expression, choosing the right transfection method and transfection reagent plays a decisive role in the success rate of transfection. This article mainly lists some methods, principles and applicability of transient transfection and stable transfection, and lists the steps and precautions of liposome cell transfection.

protocols
Protocol

Analysis And Solutions to Common Problems in Cell Culture

Experiment Tags: Cell Culture

Experiment Summary: Cell culture is a fundamental technology in biological research, but it often faces challenges that may affect experimental results. The following will analyze common problems in cell culture and provide practical solutions.

protocols
Protocol

Protocol for Cell Viability Assays: CCK-8 and MTT

Experiment Tags: CCK-8 assay, MTT assay, cell viability, drug treatment, IC50, absorbance measurement, cell seeding, aseptic technique, OD values, 96-well plate, biological assay, cytotoxicity, experimental protocol.

Experiment Summary: This protocol provides a step-by-step guide for performing CCK-8 and MTT cell viability assays, covering cell seeding, drug concentration settings, treatment duration, and absorbance measurement. It also includes important considerations for avoiding contamination, ensuring cell health, and interpreting results accurately, especially in high-throughput drug testing.

protocols
Protocol

Dual-Fluorescence Flow Cytometry for Protein Level Analysis

Experiment Tags: Dual-fluorescence, Flow Cytometry, Protein Analysis, Steady-State Protein Levels, eGFP mCherry, Transfection Efficiency, Cell Line Transfection, Protein Abundance

Experiment Summary: This protocol provides a powerful tool for studying protein expression levels under various cellular conditions, offering single-cell resolution and the ability to detect subtle differences that might be masked in bulk assays. Its versatility makes it applicable to a wide range of proteins and cellular contexts, contributing to our understanding of protein homeostasis and cellular biology.

protocols
Protocol

3D Tumorsphere Culture Protocol for Cancer Research

Experiment Tags: 3D Tumorsphere Culture, Cancer Research, Matrigel Embedding, Tumor Tissue Culture, Cell Transduction, Cryopreservation, Cell Counting

Experiment Summary: This protocol outlines a comprehensive procedure for 3D tumorsphere cultivation, including cell transduction, 3D tumorsphere culture using Matrigel, primary tumorsphere culture from fresh tumor tissue, spheroid passaging, and cryopreservation. The protocol aims to replicate the 3D environment of solid tumors, aiding researchers in studying tumor biology and conducting drug screening.

protocols
Protocol

Intraductal Injection of Adenoviral Cre for Temporally Controlled Breast Cancer Modeling in Mice

Experiment Tags: Breast Cancer, Mouse Model, Intraductal Injection, Adenoviral Cre, Temporal Control

Experiment Summary: This protocol outlines a technique for injecting adenoviral Cre (Ad-Cre) intraductally into mice mammary glands to create animal models of breast cancer. By employing cell-type-specific promoters to target mammary epithelial cells (MECs), this strategy enables fine temporal control of cancer induction and cell-type specificity. The process is described in the protocol, which includes creating and caring for floxed mice, monitoring the growth of breast tumors, and providing postoperative care.

protocols
Protocol

Protocol for Mitochondria Transfer between Mesenchymal Stem Cells and Glioblastoma Stem Cells

Experiment Tags: Mitochondria Transfer, Mesenchymal Stem Cells, Glioblastoma Stem Cells, Cell Interaction, Cell Biology.

Experiment Summary: This protocol facilitates the transfer of mitochondria from mesenchymal stem cells (MSCs) to glioblastoma stem cells (GSCs). This protocol allows for the investigation of the effects of mitochondria on GSC metabolism, proliferation, and response to therapy by isolating mitochondria from MSCs and transferring them to GSCs.

protocols
Protocol

Protocol for Rate-Controlled Cryopreservation and Thawing of Mammalian Cells

Experiment Tags: Biological Techniques, Cryopreservation, Cooling, Freezing

Experiment Summary: In this protocol, we describe the general procedure for cryopreservation of cultured cells.

* For research use only. Not intended for any clinical use.
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