Immunofluorescence analysis of Apolipoprotein A-I 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 anti-Apolipoprotein A-I (2G4) Mouse Monoclonal Antibody (LFMA0127) at 1:250 dilution 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 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.<br />
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified, human apolipoprotein A-I protein.|
|Purification||Ammonium sulfate precipitation|
|Storage buffer||HEPES with 0.15M NaCl, 0.01% BSA, 50% glycerol|
|Contains||0.03% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||Assay dependent|
|Western Blot (WB)||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.
A suggested positive control for this product is purified human Apolipoprotein A-I protein.
Apolipoprotien A-I (Apo AI) is one of the major protein components in high density lipoprotein (HDL). The molecular weight is reported to be 28.3 kDa. Apo A-I acts as a cofactor for lecithin cholesterol acyl transferase (LCAT), which is important in removing excess cholesterol from tissues and incorporating it into HDL for reverse transport to the liver. Some of the apolipoproteins act as ligands to tissue receptors, while others activate or inhibit enzymes involved in metabolic steps in the circulation or tissues. Apo A-I plays important roles in atherosclerosis, tangier disease, coronary heart disease.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.