- NCBI Gene Id:
- Protein Name:
- Glycoprotein hormones alpha chain
- Alias Symbols:
- HCG, LHA, FSHA, GPHa, TSHA, GPHA1, CG-ALPHA, CGA
- Description of Target:
- MOUSE ANTI HUMAN FSH ALPHA
- Protein Size (# AA):
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express CGA.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express CGA.
- The immunogen for anti-CGA antibody: native
- Predicted Species Reactivity:
- Predicted Homology Based on Immunogen Sequence:
- Product Format:
- Purified IgG - liquid
- Reconstitution and Storage:
- Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
- Printable datasheet for anti-CGA antibody - OASA02822
- F1 (BGN/F62/01)
- Application Info:
- ELISA : This product is suitable for use as a detection antibody in a sandwich ELISA assay with OASA02821 as the capture antibody.
- Preparation: Purified IgG prepared by affinity chromatography on Protein G
Preservative Stabilisers: 0.09% - Sodium Azide (NaN3)
- Approx Protein Conc: IgG concentration 1.0 mg/ml
Buffer Solutions: Phosphate buffered saline, pH 7.2
- Protocol Information:
- Citation: 1: Revill K, Dudley KJ, Clayton RN, McNicol AM, Farrell WE. Loss of neuronatin expression is associated with promoter hypermethylation in pituitary adenoma. Endocr Relat Cancer. 2009 Jun;16(2):537-48. Epub 2009 Feb 13. PubMed PMID:19218280.
Species: Human, Mouse
Experiment Name: IHC staining
Experiment Background: The imprinted gene, neuronatin (NNAT), is one of the most abundant transcripts in the pituitary and is thought to be involved in the development and maturation of this gland. In a recent whole-genome approach, exploiting a pituitary tumour cell line, we identified hypermethylation associated loss of NNAT. In this report, we determined the expression pattern of NNAT in individual cell types of the normal gland and within each of the different pituitary adenoma subtypes. In addition, we determined associations between expression and CpG island methylation and used colony forming efficiency assays (CFE) to gain further insight into the tumour-suppressor function of this gene. Immunohistochemical (IHC) co-localization studies of normal pituitaries showed that each of the hormone secreting cells (GH, PRL, ACTH, FSH and TSH) expressed NNAT. However, 33 out of 47 adenomas comprising, 11 somatotrophinomas, 10 prolactinomas, 12 corticotrophinomas and 14 non-functioning tumours, irrespective of subtype failed to express either NNAT transcript or protein as determined by quantitative real-time RT-PCR and IHC respectively. In normal pituitaries and adenomas that expressed NNAT the promoter-associated CpG island showed characteristics of an imprinted gene where w50% of molecules were densely methylated. However, in the majority of adenomas that showed loss or significantly reduced expression of NNAT, relative to normal pituitaries, the gene-associated CpG island showed significantly increased methylation. Induced expression of NNAT in transfected AtT-20 cells significantly reduced CFE. Collectively, these findings point to an important role for NNAT in the pituitary and perhaps tumour development in this gland.
Experimental Steps: To identify specific cell types within normal pituitary, sections were stained with antibodies directed against each of the major hormones secreted by this gland. Thus, as previously described (Simpson et al. 1999) with minor modifications, slides of formalin fixed, wax embedded normal human pituitary tissue were first re-hydrated and washed, serum blocked for 15 min at room temperature then incubated at room temperature for 30 min with antibodies directed against ACTH (mouse monoclonal, 1:2000, DAKO Cytomation, Ely, UK), GH (rabbit polyclonal, 1:6000, DAKO), FSH (mouse monoclonal, 1:5000, , Oxford, UK), PRL (mouse monoclonal, 1:2000, ) or TSH (mouse monoclonal, 1:20 000, ). Sections were then washed and incubated with biotinylated secondary antibody (Vector, Southgate, UK) for 30 min at room temperature. Antibodies were detected using the avidin–biotin complex (Vector) method using vector blue-stain as the chromogen.To observe co-localization of NNAT with each pituitary hormone, the slides were then taken through an antigen retrieval step, serum blocked for 15 min at room temperature and then incubated with an antibody directed towards NNAT (rabbit polyclonal, 1:1000, Abcam, Cambridge, UK). NNAT antibody was detected using the EnVision DAB system (DAKO Cytomation) with Dako REAL 3,3-diaminobenzidine tetrahydrochloride (DAB; DAKO Cytomation) as the chromogen. Finally, the slides were haematoxylin counterstained with a 5 min immersion in 5% copper sulphate to intensify the brown DAB stain, then de-hydrated and mounted with Pertex substitute (BDH, Lutterworth, UK) and visualized using the Nikon Eclipse e600 camera. Specificity of the antibody to NNAT was assessed by incubation with 25 mg NNAT immunizing peptide (Abcam) prior to the staining procedure described above.Co-localization of NNAT with each of the individual pituitary hormone antibodies used in this study was observed as a merging of the blue-(hormone) and the NNAT-(brown) specific stains to produce an intensity of colour that varied between dark blue and dark brown.A total of 21 adenomas, that included specimens of each of the pituitary adenoma subtypes, were stained for NNAT immunoreactivity using the EnVision DAB system as described above.
Number Of Protocols: 1
- Target Reference:
- 1. Sohn, J. et al. (2003) Orientation of follicle-stimulating hormone (FSH) subunits complexed with the FSH receptor. Beta subunit toward the N terminus of exodomain and alpha subunit to exoloop 3. J. Biol. Chem. 278: 47868-47876.