Immunofluorescent analysis of LAMP1 (green) in NRK cells. The cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS, and blocked with 1% Blocker BSA (Product # 37525), each for 15 minutes at room temperature. Cells were stained with a LAMP1 monoclonal antibody (Product # MA1-164) at a concentration of 10ug/ml in 1% Blocker BSA in PBS (right panel), or incubated in blocking buffer alone as a negative control (left panel) overnight at 4C, and then incubated with a Dylight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:1000 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI (Product # 46190). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Hamster, Human, Mouse, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Rat liver lysosomal membrane preparations|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:100-1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
MA1-164 detects glycosylated LAMP-1 at ~130kD.
M-PER (Product # 78501), IP Lysis Buffer (Product # 87787), or a 1% Triton X-100 lysis buffer are recommended for Western blot. LAMP-1 was not detected when RIPA buffer was used to generate cell lysates.
Lysosome associated membrane proteins (LAMP1), also known as Igp 120 or IgpA, is a type 1 intergral membrane protein that is transported from trans-Golgi network to endosomes and then lysosomes. Upon cell activation, LAMP1 transfer to the plasma membrane is dependent on a carboyxl-terminal tyrosine based motif (YXXI). Perturbation in the spacing between the tyrosine based motif relative to the membrane abolishes lysosome localization of LAMP1. This mutant protein then cycles between the plasma membrane and the endosome. Cell surface LAMP1 and LAMP2 have been shown to promote adhesion of human peripheral blood mononuclear cells (PBMC) to vascular endothelium, therefore they are possibly invovled in the adhesion of PBMC to the site of inflammation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Extracellular and Luminal pH Regulation by Vacuolar H+-ATPase Isoform Expression and Targeting to the Plasma Membrane and Endosomes.
MA1-164 was used in western blot to investigate luminal and extracellular pH regulation by Vacuolar H+-ATPase isoform expression and targeting to the endosomes and plasma membrane
|Smith GA,Howell GJ,Phillips C,Muench SP,Ponnambalam S,Harrison MA||The Journal of biological chemistry (291:8500)||2016|