Immunofluorescence analysis of Alpha Fetoprotein was performed using 70% confluent log phase Hep G2 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 Alpha Fetoprotein Rabbit Polyclonal Antibody (Product # PA5-16658) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A27034) 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 (Product # 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, Sheep, Pig|
|Published species reactivity||Pig, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:200|
|Western Blot (WB)||1-3 µg/ml|
* 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.
PA5-16658 targets Alpha Fetoprotein in IHC (P) and WB applications and shows reactivity with Human, Ovine, and Porcine samples.
The PA5-16658 immunogen is purified alpha-fetoprotein.
AFP is normally synthesized in the liver, intestinal tract, and yolk sac of the fetus. Antibody to AFP has been shown to be useful in detecting hepatocellular carcinomas (HCC) and germ cell neoplasms, especially yolk sac tumors.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Culture of mouse amniotic fluid-derived cells on irradiated STO feeders results in the generation of primitive endoderm cell lines capable of self-renewal in vitro.
PA5-16658 was used in immunohistochemistry to study the generation of primitive endoderm cells with self-renewal capacity following the culture of cells derived from murine amniotic fluid on irradiated STO fibroblast feeder layers
|Babic AM,Jang S,Nicolov E,Voicu H,Luckey CJ||Cells, tissues, organs (198:111)||2013|
Efficient generation of human iPSCs by a synthetic self-replicative RNA.
PA5-16658 was used in immunohistochemistry to develop a synthetic self-replicating RNA replicon expressing reprogramming factors that enables the efficient generation of induced pluripotent stem cells
|Yoshioka N,Gros E,Li HR,Kumar S,Deacon DC,Maron C,Muotri AR,Chi NC,Fu XD,Yu BD,Dowdy SF||Cell stem cell (13:246)||2013|
Differential proteome and transcriptome analysis of porcine skeletal muscle during development.
PA5-16658 was used in western blot to study developmental changes in pig skeletal muscle at the transcriptomic and proteomic levels
|Xu Y,Qian H,Feng X,Xiong Y,Lei M,Ren Z,Zuo B,Xu D,Ma Y,Yuan H||Journal of proteomics (75:2093)||2012|