The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesIHC: This antibody may be diluted to a titer of 1:100-1:500 in an ABC method.
Staining Protocol: We suggest an incubation period of 30-60 minutes at room temperature.
However, depending upon the fixation conditions and the staining system employed, optimal incubation conditions and antibody dilutions should be determined by the user.
Although not required, proteolytic treatment may enhance the immunostaining in overfixed tissues.
RelevanceHuman chorionic gonadotropin (hCG) is a glycoprotein hormone produced by trophoblastic cells of the placenta beginning 10 to 12 days after conception. Maintenance of the fetus in the first trimester of pregnancy requires the production of hCG, which binds to the corpus luteum of the ovary which is stimulated to produce progesterone which in turn maintains the secretory endometrium. hCG is present only in trace amounts in non pregnant urine and sera. It rises sharply during pregnancy. HCG is composed of two non identical, non covalently linked polypeptide chains designated as the a and b subunits. The a subunit of HCG is nearly identical to that of thyroid stimulating hormone (TSH), follicle stimulating hormone (FSH) and luteinizing hormone (LH). A germ cell tumor which is positive for cytokeratin, placental alkaline phosphatase (PLAP) and HCG but negative for EMA and AFP is probably a choriocarcinoma.
Human normal placenta. Staining is localised extracellular and in the cytoplasm. Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for rabbit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
ab9376 staining hCG in formalin-fixed, paraffin-embedded Human placenta by Immunohistochemistry.
References for Anti-hCG antibody (ab9376)
This product has been referenced in:
Wang JM et al. The human leukocyte antigen G promotes trophoblast fusion and ß-hCG production through the Erk1/2 pathway in human choriocarcinoma cell lines. Biochem Biophys Res Commun434:460-5 (2013).
Read more (PubMed: 23583402) »
Dukic-Stefanovic S et al. Chorionic gonadotropin and its receptor are both expressed in human retina, possible implications in normal and pathological conditions. PLoS One7:e52567 (2012).
Read more (PubMed: 23285091) »