Mouse Growth Hormone Receptor HEK293 Overexpression Lysate 产品信息
Product Description
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-M03H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the extracellular domain (Met 1-Gln 273) of mouse GHR (NP_034414.2) precursor was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant mouse GHR/Fc is a disulfide-linked homodimer after removal of the signal peptide. The reduced monomer consists of 497 amino acids and has a predicted molecular mass of 56.8 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rmGHR/Fc monomer is approximately 70-80 kDa due to glycosylation.
Mouse Growth Hormone Receptor HEK293 Overexpression Lysate Usage Guide
Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Mouse Growth Hormone Receptor HEK293 Overexpression Lysate Alternative Names
Mouse GHBP Overexpression Lysate;Mouse GHR/BP Overexpression Lysate
Growth Hormone Receptor Background Information
Growth hormone receptor, also known as GH receptor and GHR, is a single-pass type I membrane protein which belongs to the type I cytokine receptor family and type 1 subfamily. GHR contains one fibronectin type-III domain. Growth hormone receptor / GHR is expressed in various tissues with high expression in liver and skeletal muscle. Isoform 4 of GHR is predominantly expressed in kidney, bladder, adrenal gland and brain stem. Isoform 1 expression of GHR in placenta is predominant in chorion and decidua. Isoform 4 is highly expressed in placental villi. Isoform 2 of 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. Isoform 2 of 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.
Full Name
growth hormone receptor
References
Leung DW. et al., 1987, Nature. 330:537-43. Sobrier M-L. et al., 1997, J Clin Endocrinol Metab. 82: 435-7. Enberg B. et al., 2000, Eur J Endocrinol. 143: 71-6. Pantel J. et al., 2000, J Biol Chem. 275: 18664-9. Jorge AAL. et al., 2004, Clin Endocrinol. (Oxf.) 60: 36-40.
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