Immunofluorescent analysis of alpha-fetoprotein (green) in the endoderm derived from human ES cells. Embryoid bodies (EBs) were generated from the H9 embryonic stem cell line (WiCell Research Institute, WA09) using Gibco® KnockOut™ Serum Replacement (http://www.lifetechnologies.com/us/en/home/references/protocols/cell-culture/stem-cell-protocols/ipsc-protocols/embryoid-bodies-from-ipscs.html). After four days in suspension culture, EBs were plated on Geltrex™-coated tissue culture-treated polystyrene plates and continuously cultured for 21 days. EB cultures were then fixed and permeabilized according to the 3-Germ Layer Immunocytochemistry Kit (Product # A25538) and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with anti-alpha-fetoprotein (Product # MIA1305, 1:200 dilution, 5 ug/mL final concentration) at 4°C overnight, and then incubated with a Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:1000 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Stained cells were imaged at 40X using the EVOS® FL Auto Imaging System.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified AFP from Cord Blood|
|Storage buffer||PBS, pH 7.4|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunocytochemistry (ICC)||5 µg/ml|
|Immunofluorescence (IF)||5 µg/ml|
|Immunoprecipitation (IP)||Assay Dependent|
|Radioimmune Assays (RIA)||Assay Dependent|
|Western Blot (WB)||1 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.
MIA1305 can be used for immunofluorescence analysis of AFP in the endoderm derived from human embryonic stem cells. By Western blot, MIA1305 detects endogenous AFP protein in the early hepatocyte-like cells derived from human embryonic stem cells.
This gene encodes alpha-fetoprotein, a major plasma protein produced by the yolk sac and the liver during fetal life. Alpha-fetoprotein expression in adults is often associated with hepatoma or teratoma. However, hereditary persistance of alpha-fetoprotein may also be found in individuals with no obvious pathology. The protein is thought to be the fetal counterpart of serum albumin, and the alpha-fetoProtein A column chromatographynd albumin genes are present in tandem in the same transcriptional orientation on chromosome 4. Alpha-fetoprotein is found in monomeric as well as dimeric and trimeric forms, and binds copper, nickel, fatty acids and bilirubin. The level of alpha-fetoprotein in amniotic fluid is used to measure renal loss of protein to screen for spina bifida and anencephaly.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.