This Mouse Growth Hormone Receptor overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Growth Hormone Receptor protein (Cat: 50043-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of mouse GHR (NP_034414.2) (Met 1-Gln 273) was fused with a polyhistidine tag at the C-terminus.
The recombinant mouse GHR consists of 260 amino acids and has a predicted molecular mass of 30.3 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rmGHR is approximately 40-45 kDa due to glycosylation.
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Growth hormone receptor, also known as GH receptor and GHR, is a single-pass type I membrane protein which belongs to thetype I cytokine receptor family and type 1 subfamily. GHR contains onefibronectin type-III domain. Growth hormone receptor / GHR is expressed in various tissues with high expression in liver and skeletal muscle. Isoform4of GHR is predominantly expressed in kidney, bladder, adrenal gland and brain stem. Isoform1 expression of GHR in placenta is predominant in chorion and decidua. Isoform4is highly expressed in placental villi. Isoform2of GHR is expressed in lung, stomach and muscle. Growth hormone receptor / GHR is a receptor for pituitary gland growth hormone. It is involved in regulating postnatal body growth. On ligand binding, it couples to the JAK2 / STAT5 pathway. Isoform2of GHR up-regulates the production of GHBP and acts as a negative inhibitor of GH signaling. Defects in GHR are a cause of Laron syndrome (LARS) which is a severe form of growth hormone insensitivity characterized by growth impairment, short stature, dysfunctional growth hormone receptor, and failure to generate insulin-like growth factor I in response to growth hormone. Defects in GHR may also be a cause of idiopathic short stature autosomal (ISSA) which is defined by a subnormal rate of growth.