Apr 06, 2026
- 1Icahn School of Medicine at Mount Sinai
Protocol Citation: Elena Coccia 2026. SAM ELISA. protocols.io https://dx.doi.org/10.17504/protocols.io.n2bvjkmywgk5/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 06, 2026
Last Modified: April 06, 2026
Protocol Integer ID: 314563
Keywords: ELISA, cell lysate, protein quantification, immunoassay, sam elisa this protocol, linked immunosorbent assay, immunosorbent assay, specific proteins in the sample, sam elisa, preparing cell lysate, specific protein, elisa, assay, enzyme
Disclaimer
This protocol was generated by AI
Abstract
This protocol outlines the steps for preparing cell lysates and conducting an enzyme-linked immunosorbent assay (ELISA) to quantify specific proteins in the samples.
Materials
96-well microtiter plate (Flat-bottom polystyrene, suitable for ELISA) - 1 plate
Micropipettes (Adjustable volume (1-1000 µL)) - 1 each
Multichannel pipette (12-channel, 20-200 µL) - 1
Centrifuge (Refrigerated, capable of 10,000 x g) - 1
Sonicator (Capable of homogenizing cell pellets) - 1
Orbital shaker (Adjustable speed, for incubation) - 1
Absorbance spectrophotometer (Capable of reading at 450 nm) - 1
Reagents:
Buffer PBS - Phosphate-buffered saline for cell lysis
Antibody Anti-SAM Antibody - Specific antibody for SAM detection
Conjugate Secondary Antibody HRP Conjugate - Enzyme-linked secondary antibody for detection
Substrate TMB (3,3',5,5'-Tetramethylbenzidine) - Chromogenic substrate for HRP
Stop Solution 1M H2SO4 - Stops the enzymatic reaction
Wash Buffer 1X Wash Buffer - Buffer for washing wells
Troubleshooting
Problem
High background signal
Solution
Increase wash steps or optimize antibody dilutions.
Problem
Low signal
Solution
Ensure proper incubation times and concentrations of antibodies.
Safety warnings
Handle all reagents and samples according to safety data sheets. Use appropriate personal protective equipment (PPE) including gloves and lab coats. Dispose of all waste according to institutional guidelines.
Cell Lysate Preparation
Collect and prepare cell lysates for analysis.
Cell Collection
Centrifuge cell culture at 2,000 x g for 10 min at 4 °C.
Cell Lysis
Resuspend the cell pellet in 1-2 mL of cold PBS on ice. Sonicate the suspension for 10-15 seconds, ensuring not to overheat. Keep the sample on ice to prevent protein degradation.
Centrifugation
Centrifuge the lysate at 10,000 x g for 15 min at 4 °C. Carefully collect the supernatant without disturbing the pellet.
Storage
Aliquot the supernatant and store on ice for immediate use or at -80 °C for long-term storage.. Avoid repeated freeze-thaw cycles.
Protocol references