Immunofluorescence analysis of APOE was performed using 70% confluent log phase HepG2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with APOE Mouse Monoclonal Antibody (MA516146) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Synthetic peptide made to an internal region of human ApoE4 (within residues 100-150).|
|Storage buffer||tris glycine with 0.15M NaCl|
|Contains||0.05% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1:1000-1:2000|
* 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.
ApoE4 is one of the three major isoforms of ApoE (Apolipoprotein E). These three major isoforms (ApoE2, ApoE3, and ApoE4) correspond to three common ApoE alleles and are recognized according to their relative position after isoelectric focusing. Individuals carrying the common ApoE4 variant are at higher risk of coronary artery disease. ApoE4 is also the largest known genetic risk factor for late-onset Alzheimer's Disease (AD).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.