On-cell western [74 ] was performed with U2OS MYC-ER cells. 72,000 cells were plated in each well of a 24-well plate, and 24 hours later, treated ± 4OHT for 48 hours. Cells were then fixed with 3.7% paraformaldehyde (Sigma) but not permeabilized. Wells were washed with tris-buffered saline, and stained with primary antibodies mouse anti-LAT1 BU53 (Novus NBP2-50465AF647) or mouse IgG2A isotype control (Novus IC003R), and secondary antibodies goat anti-mouse Alexa Fluor 790 (Thermo Scientific A11357) or CellTag 700 Stain (Licor 926–41090), which is used to quantify total cell number and intensity. The stained plate was then digitally imaged using a Licor Odyssey CLx infrared imager (Licor), and well intensities in both channels (700 nM for CellTag, 800 for LAT1 or isotype control) were quantified using ImageStudio software (Licor). Individual wells were treated as biological replicates, and results are representative of two separate experiments.
Mouse igg2a isotype control
Manufactured by Novus Biologicals
The Mouse IgG2A isotype control is a laboratory reagent used as a control for immunological assays. It serves as a reference to differentiate specific antigen-antibody interactions from non-specific binding. The core function of this product is to provide a standardized control for the reliable interpretation of experimental results.
Automatically generated - may contain errors
Lab products found in correlation
Celltag 700 stain, by LI COR (2 mentions)
Mouse anti lat1 bu53, by Novus Biologicals (2 mentions)
Clx infrared imager, by LI COR (2 mentions)
Image studio software, by LI COR (2 mentions)
Paraformaldehyde (pfa), by Merck Group (2 mentions)
Goat anti mouse alexa fluor 790, by Thermo Fisher Scientific (1 mentions)
2 protocols using mouse igg2a isotype control
1
Quantifying LAT1 expression in MYC-ER cells
2
Quantifying LAT1 expression in MYC-ER cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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On-cell western [74 ] was performed with U2OS MYC-ER cells. 72,000 cells were plated in each well of a 24-well plate, and 24 hours later, treated ± 4OHT for 48 hours. Cells were then fixed with 3.7% paraformaldehyde (Sigma) but not permeabilized. Wells were washed with tris-buffered saline, and stained with primary antibodies mouse anti-LAT1 BU53 (Novus NBP2–50465AF647) or mouse IgG2A isotype control (Novus IC003R), and secondary antibodies goat anti-mouse Alexa Flour 790 (Thermo Scientific A11357) or CellTag 700 Stain (Licor 926–41090), which is used to quantify total cell number and intensity. The stained plate was then digitally imaged using a Licor Odyssey CLx infrared imager (Licor), and well intensities in both channels (700 nM for CellTag, 800 for LAT1 or isotype control) were quantified using ImageStudio software (Licor). Individual wells were treated as biological replicates, and results are representative of two separate experiments.
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