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Protocols

protocols
Protocol

What is the ELISpot Assay? Complete Guide to Principle, Steps, and Uses

Experiment Tags: ELISpot Assay, Cytokine Detection, Single-Cell Immunoassay

Experiment Summary: The ELISpot (Enzyme-Linked ImmunoSpot) assay is a highly sensitive immunological technique that detects secreted proteins—such as cytokines or antibodies—at the single-cell level. It quantifies the frequency of secreting cells, offering functional insights that conventional ELISA cannot provide.

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

E. coli Expression Experimental Protocol

Experiment Tags: E. coli Expression, Cloning, plasmid preparation

Experiment Summary: Recombinant protein expression in Escherichia coli (E. coli) is widely used in molecular biology and biotechnology due to its ease, cost-effectiveness, and rapid growth. This E. coli expression protocol outlines a standardized workflow for cloning, transformation, protein expression, and purification in the E. coli system, ensuring reproducibility and efficiency.

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

Plasmid Purification And Troubleshooting

Experiment Tags: Plasmid purification, Plasmid purification

Experiment Summary: Plasmids are small circular deoxyribonucleic acids (DNA) outside the chromosomes that replicate independently of chromosomal DNA. Although budding yeast and fission yeast can retain plasmids, the host of plasmids is almost always bacteria. This small circular DNA is widely used as a DNA vector in the fields of molecular biology, biochemistry, biotechnology, cell biology, etc.

protocols
Protocol

Comprehensive Guide to Stable Lentiviral Cell Line Construction

Experiment Tags: Lentiviral Vectors, Stable Cell Line, Gene Expression, Viral Transduction, Cell Culture, Puromycin Selection, Virus Packaging, Mammalian Cells, Transgene Expression

Experiment Summary: This protocol details the generation of stable cell lines using lentiviral vectors. The method covers plasmid preparation, 293T cell transfection for viral packaging, PEG8000-based virus concentration, target cell infection, antibiotic selection parameters, and single-clone isolation. Includes MOI calculation guidance, troubleshooting for common challenges like low viral titer and poor infection efficiency, and validation strategies for ensuring stable transgene expression.

protocols
Protocol

Bacterial Transformation Protocol

Experiment Tags: Bacterial Transformation, Bacterial Transformation Protocol, transformation, bacterial cells

Experiment Summary: Bacterial transformation is a fundamental process in molecular biology, enabling the introduction of foreign DNA into bacterial cells. Primarily used for the amplification and storage of plasmids, bacterial transformation is critical for cloning experiments.

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.

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