To make 0.5% agar gels, 2 ml 2× RPMI (made from Powder) (Sigma) was mixed with 200 μl human serum (One Lambda) and 800 μl ultrapure H2O. This was prewarmed to 37 °C in a water bath. About 2% agar was dissolved in ultrapure H2O by bringing it to a boil in the microwave and mixing it on high speed on a vortex mixer for 20 s. This process was repeated four times. One milliliter of the agar solution was added to the prewarmed mixture to make a 0.5% agar medium solution. Of the solution, 800 μl was added to each well of a 4-well 1.5 polymer tissue culture–treated chambered coverslip (Ibidi) that was precoated with 20% human serum in RPMI for 30 min at 37 °C.
Human serum
Manufactured by Ibidi
Sourced in Germany
Human serum is a complex biological fluid derived from the blood of human donors. It contains a diverse array of proteins, electrolytes, hormones, and other biomolecules that are essential for various physiological processes. This product is commonly used in cell culture applications and as a reference material in diagnostic assays. The core function of human serum is to provide a natural, protein-rich environment for the maintenance and growth of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Human serum, by Thermo Fisher Scientific (3 mentions)
Roswell park memorial institute (rpmi), by Ibidi (3 mentions)
1.5 polymer tissue culture treated chambered coverslip, by Ibidi (2 mentions)
Ccl19, by Thermo Fisher Scientific (2 mentions)
Roswell park memorial institute (rpmi), by Merck Group (1 mentions)
3 protocols using human serum
1
Preparing 0.5% Agar Gel Samples
2
Agar-Based Microenvironment for Chemoattractant Studies
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To make 0.5% agar gels, 2 mL 2x RPMI (made from powder) (Sigma St. Louis, Missouri, USA) was mixed with 200 µl human serum (One Lambda, Los Angles, California, USA) and 800 µl ultrapure H2O. This was prewarmed to 37°C in a water bath. 2% agar was dissolved in ultrapure H2O by bringing it to a boil in the microwave and mixing it on high speed on a vortex mixer for 20 seconds. This process was repeated four times. One mL of the agar solution was added to the prewarmed mixture to make a 0.5% agar medium solution. Of the solution, 800 µl was added to each well of a 4 well 1.5 polymer tissue culture treated chambered coverslip (Ibidi, Martinsried, Germany) that was precoated with 20% human serum in RPMI for 30 min at 37°C. To generate a uniform concentration of CCL19 (PeproTech), CCL19 was added to a final concentration of 100 ng mL-1 before letting the agar solidify. The agar was left to set for 1 hour. Next, a three-pronged bespoke autopsy punch was used to create a line of three wells, each of a three mm diameter and 2 mm apart in the agar (Supplementary Figure 2
3
Fabrication of 0.5% Agar Gels
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To make 0.5% agar gels, 2 mL 2x RPMI (made from Powder) (Sigma St. Louis, Missouri, USA) was mixed with 200 µl human serum (One Lambda, Los Angles, California, USA) and 800 µl ultrapure H2O. This was prewarmed to 37°C in a water bath. 2% agar was dissolved in ultrapure H2O by bringing it to a boil in the microwave and mixing it on high speed on a vortex mixer for 20 seconds. This process was repeated four times. One mL of the agar solution was added to the prewarmed mixture to make a 0.5% agar medium solution. Of the solution, 800 µl was added to each well of a 4 well 1.5 polymer tissue culture treated chambered coverslip (Ibidi, Martinsried, Germany) that was precoated with 20% human serum in RPMI for 30 min at 37°C. To generate a uniform concentration of CCL19 (PeproTech), CCL19 was added to a final concentration of 100 ng mL -1 before letting the agar solidify. The agar was left to set for 1 hour. Next, a three-pronged bespoke autopsy punch was used to create a line of three wells, each of a three mm diameter and 2 mm apart in the agar (Supplementary Figure 1).
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