295,00 995,00 

Delivery Time: 3-5-business-days

Product details

Synonyms = nuclear receptor subfamily 3 group C member 1 , GCCR , GCR , GCRST , GR , GRL

Antibody type = Recombinant Rabbit monoclonal / IgG

Clone = HMV-304

Positive control = Mature placenta: A strong GR staining should be seen in stroma cells while trophoblastic cells remain negative.

Negative control = Mature placenta: GR staining should be absent in trophoblastic cells.

Cellular localization = Nucleus

Reactivity = Human

 

 

Application = Immunohistochemistry
Dilution = 1:100 – 1:200
Intended Use = Research Use Only

Relevance of Antibody

Glucocorticoid Receptor is a pivotal multifunctional nuclear receptor protein.



Biology Behind

The glucocorticoid receptor (GR, or GCR) is a 777-amino acid multidomain nuclear protein which is coded by the NR3C1 (nuclear receptor subfamily 3, group C, member 1) gene at 5q31 which consists of ten exons (1 to 9β). Together with the other steroid receptors progesterone receptor (PR), androgen receptor (AR), the estrogen receptors (ERα and ERβ), and the mineralocorticoid receptor (MR), GR builds the nuclear receptor (NR) superfamily subgroup 3. GR is the receptor for cortisol and other glucocorticoids. It is expressed in most human cell types and regulates the transcription of thousands of genes involved in metabolism, development, stress and inflammatory responses. In the absence of its ligand, GR is bound to heat shock proteins in the cytoplasm. Ligand binding results in GR dislocation to the nucleus, binding to specific glucocorticoid response elements (GREs), recruitment of context specific transcriptional coregulators, and consequently, either activation or repression of the respective target genes. GR coregulators include a large and diverse group of proteins such as histone modifying enzymes, histone acetyltransferases or deacetylases, and many others. GR and glucocorticoids have a critical impact on a very broad spectrum of physiological processes. For example, GR appears to be an important adaptor for endocrine influence – specifically the stress response – on the brain and may play a critical role in psychological disorders such as depression and post-traumatic stress disorder. GR expression also plays a complex role in tumorigenesis which goes beyond its effects on the immune system. In several tumor types, both an oncogenic and a tumor suppressive function of GR has been found depending on specific tumor conditions.

Staining Pattern in Normal Tissues

Images describing the Glucocorticoid Receptor staining pattern in normal tissues obtained by the antibody HMV-304 are shown in our “Normal Tissue Gallery”.

Brain Cerebrum Moderate GR positivity of neuronal cells.
Cerebellum Absence of unequivocal GR staining (perhaps due to overfixation of tissue)
Endocrine Tissues Thyroid Moderate to strong GR positivity of follicular epithelial cells.
Parathyroid Moderate to strong GR positivity of epithelial cells.
Adrenal gland Moderate to strong GR positivity of medullary cells while adrenocortical cells are largely GR negative.
Pituitary gland Moderate to strong GR positivity of epithelial cells. Weak GR positivity of pituicytes.
Respiratory system Respiratory epithelium Moderate to strong GR positivity of respiratory epithelial cells.
Lung Strong GR positivity of pneumocytes.
Gastrointestinal Tract Salivary glands Moderate to strong GR positivity of all epithelial cells.
Esophagus Intense GR staining of squamous epithelial cells. Staining intensity decreases sligthly towards the surface.
Stomach derate GR positivity of superficial epithelial cells. Staining is weakest in gastric glands.
Duodenum Strong GR positivity of superficial epithelial cells. Staining is somewhat weaker in the crypts.Moderate GR positivity of Brunner gland cells.
Small intestine Moderate GR positivity of epithelial cells. GR staining is stronger in lymphocytes.
Appendix Moderate GR positivity of superficial epithelial cells. Epithelial cell staining decreases markedly towards the base of crypts where it can be absent in a fraction of cells. GR staining is markedly stronger in lymphocytes.
Colon Moderate GR positivity of superficial epithelial cells. Epithelial cell staining decreases markedly towards the base of crypts where it can be absent in a fraction of cells. GR staining is stronger in lymphocytes.
Rectum Moderate GR positivity of superficial epithelial cells. Epithelial cell staining decreases markedly towards the base of crypts where it can be absent in a fraction of cells.  GR staining is markedly stronger in lymphocytes.
Liver Moderate GR positivity of hepatocytes, ductal cells and of lymphocytes.
Gallbladder Strong GR positivity of epithelial cells.
Pancreas Moderate to strong GR positivity of acinar cells while islet cells show a strong staining.
Genitourinary Kidney Moderate to strong GR positivity of all epithelial cells.
Urothelium Weak to moderate GR positivity of the basal and suprabasal cell layers of the urohelium. Superficial cell layers are largely GR negative.
Male genital Prostate Moderate to strong GR positivity of epithelial cells.
Seminal vesicles Moderate to strong GR positivity of all epithelial cells.
Testis Intense GR positivity of Sertoli and Leydig cells. Cells of the spermatogenesis do not show significant GR staining.
Epididymis Moderate to strong GR positivity of all epithelial cells of the caput epididymis.
Female genital Breast Moderate to strong GR positivity of myoepithelial and luminal cells.s
Uterus, myometrium Strong GR positivity of myometrial cells.
Uterus, ectocervix Significant GR staining of squamous epithelial cells. Staining intensity decreases sligthly towards the surface.
Uterus endocervix Moderate GR positivity of epithelial cells.
Uterus, endometrium Moderate GR positivity of stroma cells while endometrium cells are largely GR negative.
Fallopian Tube Strong GR positivity of all epithelial cells.
Ovary Strong GR positivity of stromal cells.

Moderate to strong GR positivity of corpus luteum cells. Granulosa cells are largely GR negative.

Placenta early Strong GR staining of trophoblast and stroma cells.
Placenta mature Strong GR staining of stroma cells while trophoblast cells are negative. Moderate GR positivity of decidua cells.
Amnion Strong GR positivity of amnion cells.
Chorion Strong GR positivity of chorion cells.
Skin Epidermis Intense GR positivity of squamous epithelial cells. Staining intensity decreases slightly towards the surface.
Sebaceous glands
Muscle/connective tissue Heart muscle Strong GR positivity of heart muscle cells.
Skeletal muscle Strong GR positivity of skeletal muscle cells.
Smooth muscle Moderate to strong GR positivity of smooth muscle cells.
Vessel walls Moderate to strong GR positivity of spindle shaped cells in the aortic media.
Fat Strong GR positivity of fat cells.
Stroma Moderate to strong GR positivity of all cells.
Endothelium Moderate GR positivity of endothelial cells.
Bone marrow/lymphoid tissue Bone marrow Strong GR positivity of almost all hematopoetic cells.
Lymph node Strong GR positivity of lymphocytic cells.
Spleen Strong GR positivity of lymphocytic cells.
Thymus Strong GR positivity of lymphocytic cells. GR staining is only weak in squamous epithelial cells of corpuscles of Hassall‘s.
Tonsil Strong GR positivity of lymphocytic cells.Intense GR positivity of squamous epithelial cells with somewhat decreasing intensity towards the surface.
Remarks The vast majority of cell types show strong GR staining. Only few cells are either GR negative or show greatly reduced GR staining.

In normal tissues, GR is present in every organ and in most individual cell types. These findings are largely consistent with the RNA data described in the Human Protein Atlas (Tissue expression GR). also describing ubiquitous GR expression.

 

Positive control = Mature placenta: A strong GR staining should be seen in stroma cells while trophoblastic cells remain negative.

Negative control = Mature placenta: GR staining should be absent in trophoblastic cells.

 

Normal tissue gallery

Staining Pattern in Relevant Tumor Types

GR expression occurs in virtually all tumor entities in a large fraction of cases.

The TCGA findings on GR RNA expression in different tumor categories have been summarized in the Human Protein Atlas.

 

 

Cancer tissue gallery

Compatibility of Antibodies

No data available at the moment

Protocol Recommendations

IHC users have different preferences on how the stains should look like. Some prefer high staining intensity of the target stain and even accept some background. Others favor absolute specificity and lighter target stains. Factors that invariably lead to more intense staining include higher concentration of the antibody and visualization tools, longer incubation time, higher temperature during incubation, higher temperature and longer duration of the heat induced epitope retrieval (slide pretreatment). The impact of the pH during slide pretreatment has variable effects and depends on the antibody and the target protein.

 

All images and data shown here and in our image galleries are obtained by the manual protocol described below. Other protocols resulting in equivalent staining are described as well.

 

Manual protocol

Freshly cut sections should be used (less than 10 days between cutting and staining). Heat-induced antigen retrieval for 5 minutes in an autoclave at 121°C in pH 7,8 Target Retrieval Solution buffer. Apply HMV-304 at a dilution of 1:150 at 37°C for 60 minutes. Visualization of bound antibody by the EnVision Kit (Dako, Agilent) according to the manufacturer’s directions.

Potential Research Applications

The diagnostic, prognostic, and predictive relevance of GR expression in tumors and in preneoplastic disease is unknown.

Evidence for Antibody Specificity in IHC

There are two ways how the specificity of antibodies can be documented for immunohistochemistry on formalin fixed tissues. These are: 1. Comparison with a second independent method for target expression measurement across a large number of different tissue types (orthogonal strategy), and 2. Comparison with one or several independent antibodies for the same target and showing that all positive staining results are also seen with other antibodies for the same target (independent antibody strategy). 

 

Orthogonal validation: For proteins such as GR which are expressed in all organs, orthogonal validation is not suited.

Comparison of antibodies: True expression of GR in all cell types found to be GR positive by HMV-304 is corroborated by identical stainings obtained by another commercially available independent antibody (termed “validation antibody”). Both antibodies showed a significant reduction or loss of GR expression in the same cell types (upper cell layers of non-keratinizing squamous epithelium and of the urothelium, cells of the spermiogenesis, granulosa cells of the ovary, adrenocortical cells, and trophoblastic cells of the mature (but not of the first trimenon) placenta).

 

Normal tissue gallery