Immunofluorescence analysis of Apolipoprotein A5 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 Apolipoprotein A5 (1G5G9) Mouse Monoclonal Antibody (MA116809) 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.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Ni-NTA purified truncated recombinant human APOA5 expressed in E. Coli strain BL21 (DE3).|
|Contains||0.1% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:20|
|Immunohistochemistry (IHC)||5 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||5 µg/ml|
|Western Blot (WB)||1:500-1:2000|
* 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.
The target sequence has 83% sequence homology with bovine, 79% sequence homology with rat and 78% sequence homology with mouse. This antibody is not recommended for ICC/IF applications.
Apolipoprotein A5 (ApoA5) is fast gaining attention as a key regulator of serum triglyceride concentrations. An ApoA5 mouse knock-out model produced an approximately four fold increase in serum triglycerides, whereas a knock-in model with human ApoA5 produced 50-70% lower concentrations of mouse serum triglycerides. In addition, peroxisome proliferator-activated receptor-4 (PPAR-alpha) agonists, which are used clinically to lower serum triglyceride concentrations, cause increased ApoA5 mRNA expression. Recently, it was demonstrated that ApoA5 is present in human serum detected by polyclonal antibodies against both the NH2 and COOH termini, although at much lower concentration than other apolipoproteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.