Cell Signaling Technology
EEA1 () Rabbit mAb
3288
C45B10
Info:Supplier Page View Product Specs
Browse Similar Products: EEA1 Antibodies
Used for cell fractionation experiments in mouse neutrophils. Was extremely clean and reliable at IP / organelle isolation of early endosomes.
Immunoprecipitation
Mouse Neutrophils
1:1000 for WB 1:1000 for IP
5 % BSA (for WB)
1:3000 anti-rabbit HRP
na
HRP
In the image below mouse neutrophils were gently lysed by sonication. granules were spun down at 10,000x g before the early endosomes were IP'd from the supernatant by incubation for 1hr on ice in the presence of anti-EEA1 at 1:1000. Protein A magnetic beads weer then added for a further hour to bind EEA1. Remaining supernatant was spun at 200,000 xg for 1 hour to pellet the plama membrane, and the remaining supernatant was deemed cytosol. Equivilent protein was loaded onto a gel and first blotted for EEA1 (1:1000) and then Rab5 (another marker of early endosomes)
N/A
for IP we found that Ph6 buffer was essential, otherwise the EEA1 falls out of the membrane of the early endosomes and you only pull down this protein.
Great product.
Nothing
as you can see from the blots, exceptionally pure purification.