Western blot analysis of alpha Fetoprotein was performed on HepG2 whole cell lysates. Cell lysates were mixed with reducing (Lane 1) or non-reducing (Lane 2) Laemmli SDS-PAGE sample buffer and 5ul of lysate, corresponding to 6.25 x 10^5 HepG2 cells, was loaded per well on a 10% polyacrylamide gel. Proteins were transferred to a membrane and the membrane was blocked with 5% milk overnight at 4C. The membrane was probed with an alpha Fetoprotein monoclonal antibody (Product # MA1-19178) at a dilution of 1:500-1:2000 for 1 hour at RT, followed by a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:1000 for 1 hour at RT. Detection was performed using a chemiluminescent substrate
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified alpha Fetoprotein.|
|Storage buffer||PBS, pH 7.4|
|Contains||15mM sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|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 MA1-19342 anti-alpha Fetoprotein antibody (clone AFP-11) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody MA1-19178 (clone AFP-01).
Alpha Fetoprotein is 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 and 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.