Streamlining ELISA Workflows with PIPETMAN M96

Enzyme-linked immunosorbent assay (ELISA) is a widely used analytical technique for detecting and quantifying substances such as proteins, antibodies, and hormones in biological samples. Whether used in clinical diagnostics, food safety, or biomedical research, ELISA demands meticulous pipetting accuracy and consistency across microplates to ensure reliable results.

Traditionally, ELISA workflows have relied on manual pipetting, which, while effective, can be time-consuming and prone to variability. As laboratories scale up their testing capacity, the need for high-throughput, ergonomic, and reproducible pipetting solutions becomes increasingly critical.

PIPETMAN® M96: A Game-Changer for ELISA Plate Preparation

PIPETMAN M96, Gilson’s motorized 96-channel electronic pipette, is a powerful solution for ELISA workflows. Designed for high-throughput applications, it enables simultaneous dispensing across entire microplates with precision and ease. Its key features include:

• Motorized 96-channel dispensing for uniform and quick sample distribution. Performing a simultaneous deposition step simplifies the ELISA workflow and enhances assay precision.
• Touchscreen interface for intuitive protocol setup and execution
• Custom mode for saving and reusing personalized workflows for improved reproducibility
• Partial tip fitting for flexible plate usage when analyzing fewer samples
• Compatibility with standard, SLAS-format, 96- and 384-well plates and standard PIPETMAN DIAMOND Tips

These features make PIPETMAN M96 an ideal tool for laboratories performing large volumes of ELISA tests, especially in clinical and diagnostic settings.

Benefits for High-Throughput Labs

By integrating PIPETMAN M96 into ELISA workflows, laboratories can:

• Increase throughput by filling entire plates in a single step
• Reduce variability with motorized precision dispensing
• Minimize operator fatigue through ergonomic design
• Save time and resources with reusable protocols and partial plate handling. Motorized 96-channel pipetting cuts plate setup time dramatically
• Ensure reproducibility across assays and operators

Whether you're conducting pathogen detection, biomarker analysis, or routine screening, PIPETMAN M96 empowers your lab to deliver consistent, high-quality results with greater efficiency.

Executing an ELISA Workflow with PIPETMAN M96

The following protocol illustrates how PIPETMAN M96 can be used to execute key steps in an ELISA workflow. It is based on Human Immunodeficiency Virus Type 1 (HIV-1) p24 antigen detection assay using ZeptoMetrix® RETRO-TEK HIV-1 p24 Antigen ELISA kit¹:

1. Specimen Preparation – Treat specimens by adding 50 µL of Lysing Buffer into 450 µL specimen and mix well, using PIPETMAN M96 P96x200M.
2. Plate Washing (General Procedure) – Wash each well of the microplate or the selected number of strips six times by dispensing twice 150 µL of 1X Plate Wash Buffer and aspirate, using PIPETMAN M96. Thoroughly blot by striking an inverted microplate on a pad of absorbent towels. Continue striking until no droplets remain in the wells. Do not allow washed plates to dry completely prior to sample addition. Drying will adversely affect test results.
3. Standard and Blank Preparation – Leave one well of the microtiter plate empty during the assay. This well is used for a substrate blank. Pipette 200 µL of standards #1-6 into duplicate wells.
4. Sample Addition – Pipette 200 µL of each specimen, as prepared in Step 1, into duplicate wells.
5. Sample Incubation – Cover microplate with a plate sealer and incubate as follows: a. Overnight at 37°C ± 1°C, for serum or plasma specimens treated with RETRO-TEK HIV-1 ICx/CRx Kit. b. Two hours or overnight at 37°C ± 1°C, for untreated serum or plasma, culture fluids or other biological specimens.
6. Plate Washing – Aspirate and wash plate as described in Step 2.
7. Add Detector Antibody – Using PIPETMAN M96 or a PIPETMAN L multichannel P200L, 8 or 12 channels, pipette 100 µL of reconstituted HIV-1 p24 Detector Antibody into each well, except the substrate blank – simply remove one pipette tip from your device to leave the substrate blank. Cover the microplate with a sealer and incubate for 1 hour at 37°C ± 1°C.
8. Plate Washing – Aspirate and wash plate as described in Step 2.
9. Add Streptavidin-Peroxidase – Using PIPETMAN M96, pipette 100 µL of the Streptavidin-Peroxidase Working Solution into each well except the substrate blank - Do not forget to remove one tip of your device to leave the substrate blank. Cover the microplate with a sealer and incubate for 30 minutes at 37°C ± 1°C.
10. Plate Washing – Aspirate and wash plate as described in Step 2.
11. Add Substrate Solution – Using PIPETMAN M96, pipette 100 µL of Substrate Working Solution into all wells and incubate uncovered for 30 minutes at room temperature (18°- 25°C). A blue color will develop in wells containing viral antigen.
12. Stop the Reaction – Stop the reaction by simultaneously pipetting 100 µL of Stop Solution into all 96 wells with PIPETMAN M96. A color change from blue to yellow will result.
13. Result Analysis – Within 15 minutes, read the optical density of each well at 450 nm using a microplate reader.

This protocol can be programmed and saved in the Custom mode of PIPETMAN M96, allowing users to execute it repeatedly with minimal setup time. The flexibility to use partial tip fitting also enables efficient use of reagents and consumables when only a subset of wells is needed. Additionally, no costly, specific pipette tips or cartridges are required to operate this 96-channel electronic pipette.

Elevate Your ELISA Workflow

PIPETMAN M96 is more than a pipette; it’s a workflow enhancer. With its motorized 96-channel design, customizable protocol management, and ergonomic features, it brings automation and precision to ELISA plate preparation. For labs seeking to scale their operations without compromising data integrity, PIPETMAN M96 is the smart choice.

Ready to upgrade your ELISA workflow? Discover how PIPETMAN M96 can simplify your processes and boost your lab’s productivity.

Sources:

1. HIV-1 p24 Antigen ELISA Test Procedure PI0801200 rev 05 by ZeptoMetrix®