Protocols
Protocol of SARS-CoV-2 Pseudovirus (PSV)-Based Neutralization Assay For Vaccines, therapeutic antibodies, peptides and compounds against COVID-19
GeneMedi’s Pseudovirus Based Neutralization Assay (PBNA) is a conventional assay method that is suitable for High-Throughout Screening (HTS) without live virus engaged.
Preparation of AuNP conjugates with recombinant antigens, protein or antibodies for immunoassay -- Diagnostic Application Protocol
To prepare the AuNP (gold nanoparticle) conjugate, the recombinant protein dissolved in PBS (1 mg/mL) was added to the mixture of 1mL AuNP colloid (40nm in diameter, OD=1) and 0.1mL of borate buffer (0.1 M, pH 8.5). Incubation for 30 min at room temperature.
Protocol of SARS-CoV2 (2019nCoV) Spike-RBD antibodies standard biopanning and competitive biopanning
S-RBD was diluted with 0.05 M PH 9.6 carbonate buffer [1]. Then add 100 μL to each well of 96-well Maxisorp plates at 4°C overnight.
Protocol of SARS-CoV2 (2019nCoV) NP paired antibodies sandwich ELISA
The Capture Antibody #1 was diluted to a concentration of 1 ~ 10 ug/ml with 0.05 M PH 9.6 carbonate buffer. Then add 100 μL to each well of polystyrene plate at 4℃ overnight.
Detection Antibody HRP labeling For diagnostics application (ELISA)
Add 200 μL of 0.1M NalO4 solution [1] (freshly prepared just before use) to the solution obtained in step 1 and stirred for 20 minutes at room temperature. Protect from light.
Spike & ACE2-blocking activities assay
Recombinant human ACE2 was diluted to a concentration of 5μg/ml with 0.05 M PH 9.6 carbonate buffer. Then add 100 μL to each well of polystyrene plate at 4°C overnight.
Adeno-associated virus (AAV vector) - Production/Packaging Protocol, Guidelines
The gene of interest is cloned into one of the ITR/MCS-containing AAV vectors to generate AAV-GOI. The purity and RNAse contaminants of AAV viral plasmid should be taken into consideration.
T7 Endonuclease I (T7E1) Assay Protocol for CRISPR/Cas9 gRNA validation
Design genomic DNA primers that are approximatley 18 to 22 basepairs in length and have 45-55% GC content. For best results, use primers with a Tm greater than 55 C. Design primers to yield amplicon length between 400-500 bp. In addition, design primers so that the predicted cleavage site is not in the center of the amplicon and the detection reaction will yield two distinct product bands.
Protocol of CRISPR /Cas9 mediated gene knockout in vitro and in vivo
Genemedi has launched a comprehensive AAV packaging service combined with CRISPR/Cas9, the versatile genome-editing platform. The followings are some protocols of CRISPR /Cas9 mediated gene knockout in vitro and in vivo.
Adenovirus - Production/Packaging Protocol, Guidelines
The gene of interest is cloned into one of the ITR/MCS-containing adenovirus vectors to generate pAd-GOI. The purity and RNA contaminants of viral plasmid should be taken into consideration.
Lentivirus Infection Protocol for stable cell line development (CLD)
This protocol is for the stable cell line construction based on puromycin selection.
Lentivirus vector-Production/Packaging Protocol,Guidelines And References
The gene of interest is cloned into one of the LTR/MCS-containing lentivirus vectors to generate pLV-GOI. The purity and RNA contaminants of viral plasmid should be taken into consideration.
Protocol for virus injection in vivo
Animal experiment is essential in biomedical research. It contains establishment of a nude mouse tumor model and other disease model. Besides, gene transfection in vivo is included as well.
GeneMedi's protocol / procedure for the diagnostics application-immunoassay
Chemiluminescence immunoassay (CLIA) is a sensitive technique that detects specific substances in biological samples. It utilizes light emission during antigen-antibody complex formation, offering enhanced sensitivity and a broader range. CLIA finds applications in clinical diagnostics, drug testing, and forensics, playing a crucial role in modern research and healthcare.
GeneMedi's protocol / procedure for the diagnostics application-CLIA
Chemiluminescence immunoassay (CLIA) is a sensitive technique that detects specific substances in biological samples. It utilizes light emission during antigen-antibody complex formation, offering enhanced sensitivity and a broader range. CLIA finds applications in clinical diagnostics, drug testing, and forensics, playing a crucial role in modern research and healthcare.
CLASSIFICATION of Chemiluminescence Immunoassay
Chemiluminescence immunoassay techniques, such as chemiluminescence enzyme immunoassay and electrochemiluminescence immunoassay, offer sensitive and rapid detection in diagnostics. These innovative methods push the boundaries of diagnostic capabilities, allowing for improved accuracy and efficiency in medical testing.
Advantages and disadvantages of chemiluminescence immunoassay
CLIA is a highly sensitive immunoassay technique that rapidly detects multiple analytes. It has advantages like high sensitivity, but disadvantages such as high cost, complex instrumentation, and potential interference. Nevertheless, CLIA is valuable in disease diagnosis and monitoring in diverse fields.
What is Sandwich Chemiluminescence immunoassay
Sandwich chemiluminescence immunoassay is a precise diagnostic technique. It employs two antibodies in a sandwich format, allowing for increased accuracy and specificity. This method utilizes chemiluminescent signals to quantify and analyze target analytes. Its versatility makes it invaluable in biomedical research and diagnostics.
What is Electrochemiluminescence immunoassay
Electrochemiluminescence immunoassay (ECLIA) has emerged as a potent technique, combining electrochemistry and luminescence for precise analyte detection. With advantages like low interference, wide dynamic range, and enhanced signal-to-noise ratio, ECLIA holds immense promise in biomedical research and clinical diagnostics.
What is Magnetic particle-based chemiluminescence enzyme immunoassay
The magnetic particle-based chemiluminescence enzyme immunoassay (MP-CL-EIA) combines magnetic particles and a highly sensitive chemiluminescence reaction for rapid and sensitive analyte detection. With diverse applications in clinical diagnostics, environmental monitoring, and bioanalytical research, MP-CL-EIA has revolutionized the field of analytical chemistry.
GeneMedi's protocol / procedure for the diagnostics application-ELISA
ELISA (enzyme-linked immunosorbent assay) has revolutionized immunology and diagnostics. This versatile lab technique allows for the sensitive detection and quantification of proteins, antibodies, and hormones in biological samples. Its precision and specificity have led to significant advancements in medical research and diagnostic testing.
What is Direct ELISA
Direct ELISA is a straightforward and efficient technique used for the direct detection and quantification of target molecules. By eliminating the need for secondary antibodies and amplification steps, this method offers a simplified and time-saving approach without compromising sensitivity or specificity.
What is Indirect ELISA
The Indirect ELISA technique involves using a primary antibody to bind to the target molecule, followed by a secondary antibody conjugated with an enzyme. This amplifies the signal, allowing for the detection of low levels of analytes. The Indirect ELISA is a versatile and widely used method for quantifying and detecting target molecules.
What is Competitive ELISA
Competitive ELISA is a reliable method for quantifying target molecules. Labeled antigens compete with unlabeled target molecules for binding to immobilized antibodies. The resulting signal is inversely proportional to the target molecule concentration, making it a valuable tool in diagnostics and research.
What is Sandwich ELISA
Sandwich ELISA is a sensitive and specific technique that involves capturing the target molecule between two antibodies. It offers enhanced signal amplification and lower detection limits compared to other methods. This makes it well-suited for analyzing complex samples with multiple analytes, making it a valuable tool in diagnostics and research.
A summary of the advantages and disadvantages of the different ELISA techniques
In summation, the various ELISA techniques have distinct pros and cons. Direct ELISA is fast but less sensitive, while indirect ELISA offers signal amplification but takes longer. Sandwich ELISA has high sensitivity but requires specific antibody pairs, and competitive ELISA allows for quantification but demands labeled analytes.
GeneMedi's protocol / procedure for the diagnostics application-LFIA
Lateral Flow Immunoassay (LFIA) is a rapid and user-friendly diagnostic technique that utilizes capillary action to detect target analytes. Its simplicity and minimal sample preparation requirements have made LFIA a preferred choice in diverse industries, such as clinical diagnostics and food safety.
CLASSIFICATION of lateral flow tests
Lateral flow tests come in various types to suit different needs. Antigen tests detect viral or bacterial proteins, antibody tests identify specific antibodies, and nucleic acid tests detect genetic material. Choosing the right test depends on the desired level of sensitivity and specificity for the intended application.
GeneMedi's protocol / procedure for the diagnostics application-PETIA
Particle enhanced turbidimetric immunoassay (PETIA) is a popular clinical diagnostic technique that utilizes latex particles coated with antibodies. These particles form complexes with target analytes in samples, leading to increased turbidity. This allows for quick and precise quantification of analytes, making PETIA an invaluable tool in medical diagnostics.
GeneMedi's protocol / procedure for the diagnostics application-Immunonephelometry
Immunonephelometry is a powerful clinical diagnostic technique that quantifies specific analytes by measuring the intensity of light scattered by immune complexes. Utilizing a nephelometer, this method offers exceptional sensitivity and precision, making it a valuable tool in various medical applications where accurate quantification is essential.
GeneMedi's protocol / procedure for the diagnostics application-IHC
Immunohistochemistry (IHC) is a critical technique in pathology, enabling the precise detection and localization of antigens within tissue samples. By utilizing specific antibodies, IHC offers valuable insights into disease processes and guides treatment strategies by visualizing target molecules in their cellular context.
GeneMedi's protocol / procedure for the diagnostics application-FACS
Flow cytometry, also known as fluorescent activated cell sorting (FACS), is a versatile technique used for analyzing and sorting individual cells based on their characteristics. By utilizing fluorescently labeled antibodies or dyes, flow cytometry enables high-throughput and multiparametric analysis of cell populations, making it a valuable tool in various fields of research.
Antibody titration protocol using 96 well plates
An antibody titration protocol using 96 well plates is a valuable method for determining the optimal antibody concentration. By immobilizing the target antigen and testing various antibody dilutions in a high-throughput manner, researchers can accurately assess the dilution that provides reliable and specific detection, ensuring optimal experimental results.
Protocol for cell sorting
A protocol for cell sorting is essential for isolating specific cell populations. Utilizing flow cytometry technology, this protocol enables researchers to efficiently sort cells based on desired markers or characteristics. By obtaining purified and homogeneous cell populations, this protocol enhances the accuracy and reliability of downstream analysis, benefiting research outcomes.
GeneMedi's protocol / procedure for the diagnostics application-Octet system
The Octet system employs biolayer interferometry technology to measure biomolecular interactions, providing rapid and sensitive analysis of protein-protein interactions and binding affinities. Widely utilized in drug discovery and structural biology research, this advanced tool facilitates the characterization and understanding of complex biomolecular interactions.
immunoassay-summary
In conclusion, each immunoassay technique has its own strengths and weaknesses. ELISAs offer high sensitivity but can be time-consuming, while fluorescence assays provide enhanced sensitivity and multiplexing capabilities despite potential signal interference. Chemiluminescent assays excel in sensitivity but can be expensive. The choice depends on specific assay needs.
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