I tried to monitored CD107a labelling in mouse NK cells after infection (compared to uninfected) but I'm unhappy with the plot I got.
CD107a Ab clone 1D4B rat IgG2a,k from BD Bioscience
cell are incubated 5h at 37°C in incubator with 0,5uL Ab per well.
isotope (at the same concentration) is performed the same.
How do you stain your cells for CD107a in mice?
Surface staining, ICS,...?
Thanks for your help
I do 4hr incubation and stain with PE-CD107a, same clone as yours. But at the same time during incubation I have monensin present from BD at 1:1500 dilution. Presumably this helps with stabilizing the pH in the lysosomes to preserve integrity of PE.
Check paper below for more details
Amended on 22 October 2018
I have searched CDs among my human protein database (please see file; HepG2 Fucoidan and JMBT Alopecia).
I have found CD5 antigen-like/IgM-associated peptide, CD8-alpha, CD8-beta, CD13, CD14, CD30, CD40, CD44, CD45, CD58, CD61, CD88, CD98, CD99, CD107a, CD129, CD154.
Only healed hepatocyte HepG2 (cultured with fucoidan for 3 days) has CD107a/Lysosome-associated membrane glycoprotein 1/CD107 antigen-like family member A/LAMP-1 at 0.21 μg/mg of cell protein.
This upregulation may be caused by Non-structural protein 4b (Human coronavirus 229E/HCoV-229E) at 0.20 μg/mg of cell protein.
Serum of healthy female (52y) has Lysosomal-trafficking regulator/Beige homolog at 14.5 μg/mg of serum protein. This serum has Replicase polyprotein 1a (Human coronavirus 229E/HCoV-229E) at 10.3 μg/mg of serum protein.
Serum of GSD-1b patient (after 2mo of Liver tranplantation) has Lysosomal-trafficking regulator/Beige homolog at 5.3 μg/mg of serum protein. This up regulation may be caused by the effect of Transplantation.
I do not know about Mouse. But, Coronavirus-infected Mouse may have Lysosome transport-related proteins.
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