Apr 07, 2026
CXCL1 ELISA
- 1Icahn School of Medicine at Mount Sinai
Protocol Citation: Elena Coccia 2026. CXCL1 ELISA . protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gp76r1gzp/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 07, 2026
Last Modified: April 07, 2026
Protocol Integer ID: 314561
Keywords: ELISA, sample preparation, assay procedure, antigens, biological samples, cxcl1 elisa, immunosorbent assay, linked immunosorbent assay, specific antigens in various biological sample, elisa, specific antigen, assay procedure for an enzyme, various biological sample, assay, procedures for sample preparation
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Abstract
This protocol outlines the procedures for sample preparation and the assay procedure for an Enzyme-Linked Immunosorbent Assay (ELISA) to quantify specific antigens in various biological samples.
Materials
96-well microplate (High-binding capacity, suitable for ELISA) - 1 plate
Plate shaker (Adjustable speed, suitable for 400 rpm) - 1 unit
Centrifuge (Capable of 2,000 x g and 10,000 x g) - 1 unit
Pipettes (Multichannel and single-channel) - 1 each
Reservoir trough (For reagent preparation) - 1 unit
Paper towels (For drying and cleaning) - As needed
Reagents:
Sample Diluent Sample Diluent NS - Buffer for diluting samples
Wash Buffer 1X Wash Buffer PT - Buffer for washing wells
Substrate TMB Development Solution - Chromogenic substrate for detection
Stop Solution 1M Sulfuric Acid - Stops the enzymatic reaction
Antibody Cocktail Specific Antibody - Detection antibody for the target antigen
Troubleshooting
Problem
High background color
Solution
Check for nonspecific binding; optimize blocking conditions.
Problem
Low signal
Solution
Ensure proper dilution of antibodies; check incubation times.
Problem
Inconsistent results
Solution
Verify pipetting technique; ensure uniform sample preparation.
Safety warnings
Handle all reagents and samples according to safety data sheets (SDS). Wear appropriate personal protective equipment (PPE) including gloves, lab coat, and safety goggles.
Sample Preparation
Prepare samples according to the type of biological material. For cell culture supernatants: Centrifuge at 2,000 x g for 10 minutes to remove debris. Dilute in as needed in Sample Diluent NS. Store at −20 °C.
Prepare Working Standard solutions.
Assay
Equilibrate reagents at room temperature.
Use 96-well plate strips as supplied; do not rinse before adding reagents.
Add 50 µl of each sample or standard to appropriate wells.
Add 50 µl of Antibody Cocktail to each well.
Seal the plate and incubate for 1 hour at room temperature on a plate shaker at 400 rpm.
Wash each well 3 times with 350 µl 1X Wash Buffer PT. Aspirate or decant, then dispense 350 µl Wash Buffer PT into each well. Leave for at least 10 seconds per wash.
Add 100 µl of TMB Development Solution to each well.
Incubate for 10 minutes in the dark on a plate shaker at 400 rpm.
Add 100 µl of Stop Solution to each well.
Shake on a plate shaker for 1 minute to mix.
Record OD at 450 nm on a plate reader.