Department of Molecular Genetics

Description

Recombinant human GHR is an active protein expressed from mammalian cells, with a C-terminal MFC-Avi tag. Its expression region is the DNA fragment encoding amino acid residues 27-264 of the human GHR protein. The purity of this GHR protein is greater than 95% as measured by SDS-PAGE. This recombinant Hormone GHR protein migrated to the band with a molecular weight of approximately 67 kDa on the gel. Its endotoxin level is less than 1.0 EU/ug determined by the LAL method. And its bioactivity has been validated in ELISA. In functional ELISA, this Biotinylated human GHR binds to human GH1, with a constant EC50 of 2067-3208 ng/ml. This biotinylated GHR protein could be used to isolate GHR antibodies from samples for further analysis with high sensitivity. It is in stock now.

GHR, a dimeric amino acid receptor, is widely expressed on GH target cells. The GH-GHR-IGF1 axis plays important roles in somatic growth, including cell proliferation, differentiation, division, cell cycle control, immunity, and survival. Aberrations in GHR signalling have been linked to various diseases and chronic conditions such as cancer, ageing, and inflammation.

Purity: greater than 95% as determined by SDS-PAGE.

Endotoxin: Less than 1.0 EU/ug as determined by the LAL method.

Activity

Measured by its binding capacity in a functional ELISA. Immobilized human GH1 (CSB-MP009407HU) at 2 µg/ml can bind to biotinylated human GHR, the EC50 is 2.067-3.208 ng/ml.

Destination Names: GHR

Uniprot No.: P10912

Alternative Names: (GH-binding protein)(GHBP)(serum-binding protein)

Species: Homo sapiens (Human)

Source: Mammalian cell

Expression region: 27-264aa

Mole Weight: 56.6

Protein length: Partial

Tag information: C-terminal MFC-Avi-tagged

Form: Lyophilized powder

Note: We will preferably ship the format we have in stock, however, if you have any special requirements for the format, please remark your requirement when placing the order, we will prepare according to your demand.

Buffer: Lyophilized from 0.2 μm filtered PBS, 6% trehalose, pH 7.4

Reconstitution

We recommend that this vial be briefly centrifuged before opening to bring the contents to the bottom. Reconstitute protein in sterile deionized water at a concentration of 0.1-1.0 mg/mL. We recommend adding 5-50% glycerol (final concentration) and an aliquot for long-term storage at -20°C/-80°C. Our final default glycerol concentration is 50%. Customers could use it for reference.

Storage Conditions

Store at -20°C/-80°C upon receipt, need to be aliquoted for multiple uses. Avoid repeated cycles of freezing and thawing.

Shelf life

Shelf life is related to many factors, storage condition, buffer ingredients, storage temperature and the stability of the protein itself. Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C. The shelf life of the lyophilized form is 12 months at -20°C/-80°C.

Delivery time: 3-7 business days

Notes: Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Recombinant hormones

The hormone is a kind of biochemical substance produced by multicellular glands and then transported by the circulatory system to the target organ to coordinate its physiology and behaviour. Hormones function as a serious form of communication between different organs and tissues. Hormones regulate a variety of physiological and behavioural activities, as well as digestion, metabolism, respiration, tissue function, etc. Hormones deliberately affect the target tissue by binding to specific receptor proteins and causing a specific action on the target cell. When a hormone binds to the receptor protein, it results in the activation of a signal transduction mechanism.

Ultimately, this leads to cell-type-specific genomic responses that cause the hormone to activate genes that regulate protein synthesis. Hormones can be divided into two categories: water-soluble and fat-soluble. In the first category, like protein hormones and catecholamines, they are water-soluble and therefore easily transported through the circulatory system. The next category, like steroid and thyroid hormones, are fat-soluble. For their distribution, they must bind to carrier plasma glycoproteins to form ligand-protein complexes.

BiologicsCorp(BIC) mainly manufactures two types of hormones: parathyroid hormone (PTH) and exedin. Expedia is a hormone discovered from lizard venom, it plays a role in enhancing glucose-dependent insulin secretion, suppresses inappropriately elevated glucagon secretion, and slows gastric emptying in vivo, and this will be helpful in reducing the weight. PTH increases the concentration of calcium in the blood.

Cellular differentiation determines cell fate

Cell differentiation Recombinants is the process in which a cell changes from one type to many different types. In the process of cell differentiation, there are differences in morphological structure and physiological function. All organisms start from a single cell. For example, humans derive from fertilized eggs, and this process involves the differentiation of embryonic stem cells. Cellular differentiation occurs throughout life. For example, hematopoietic stem cells differentiate into various immune cells. The abnormal differentiation of cells can lead to cancer cells. Cancer cells divide indefinitely, forming tumours and endangering human health.

Cellular differentiation involves a variety of signal transduction processes:

1. MAPK signalling pathway

2. Phosphatidylinositol (PLC) signalling pathway

3. cAMP/PKA signalling pathway

4. Via JAK-STAT

5. PI3K-AKT-mTOR signalling pathway

6. Wnt signalling pathway

7. TGF-β Superfamily Signaling Pathway

1. MAPK signalling pathway

MAPK is a mitogen-activated protein kinase, a class of protein kinases with dual phosphorylation of serine and tyrosine in the cytosol. The MAPK signalling pathway activates transcription factors and regulates gene expression through a cascade reaction (MAPKKK-MAPKK-MAPK). MAPK can trigger the activation of transcription factors in the nucleus, participate in the process of signalling from the cell surface to the nucleus, and regulate cell proliferation and differentiation. Currently, there are 4 known MAPK signalling pathways, including the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK, also known as SAPK), p38, and ERK5 pathways.

1.1 ERK-MAPK signalling pathway

In the MAPK signalling pathway, the ERK pathway acts mainly through the Ras-Raf-MEK-ERK cascade. The main process of this pathway: the growth factor activates the receptor by binding to the receptor tyrosine kinase, and the activated receptor tyrosine kinase activates the Ras protein, then the Ras protein phosphorylates Raf, and the activated Raf phosphorylates the MEK waters. below. MEK can phosphorylate and activate ERK, which is transferred to the nucleus and regulates gene expression by activating other kinases or transcription factors.

The ERK-MAPK signalling pathway plays a role in the differentiation of mesenchymal stem cells (MSCs) into adipocytes. In the early stage of adipocyte differentiation, ERK1/2 promotes adipocyte differentiation by promoting the expression of C/EBPα and PPARγ. In the late stage of adipocyte differentiation, activated ERK1/2 phosphorylates PPARγ to inactivate it and inhibit adipocyte differentiation. This pathway can also affect the proliferation and differentiation of red blood cells. Studies have shown that the ERK signalling pathway is also involved in signal transduction of osteoblast differentiation and proliferation.

1.2 Via JNK-SAPK

c-Jun N-terminal kinase (JNK), also known as stress-activated protein kinase (SAPK), is another subclass of MAPK in mammals. The JNK signalling pathway can affect a variety of vital processes, such as cell growth, cell differentiation, and cell death. JNK can change the level of osteocalcin mRNA, thus JNK activation can induce osteoblast differentiation. The JNK signalling pathway also plays an important role in the regulation of adipocyte differentiation.

1.3 via p38 MAPK

The p38 signalling pathway is an important component of the MAPK family. p38 MAPK can be activated by a variety of extracellular stress responses, including ultraviolet rays, radiation, and proinflammatory factors. The p38 pathway plays a very important role in the osteogenic differentiation of mesenchymal stem cells (MSCs). Inhibition of the p38 MARK pathway downregulates the activity of protein kinase C (PKC), which plays an important role in the osteogenic differentiation of cells.

In addition, the transforming growth factor and the bone morphogenetic protein BMP-2 induce the transcriptional expression of Runx2/Cbfa1 through the p38 MAPK pathway. Among them, Runx2 is a key target gene affecting osteogenic activity, and Cbfa1 regulates MSC differentiation into osteoblasts at the transcriptional level.

2. Phosphatidylinositol (PLC) signalling pathway

In the phosphatidylinositol signalling pathway, extracellular signalling molecules bind to G protein-coupled receptors, activating phospholipase C (PLC-β) on the plasma membrane, causing phosphatidylinositol bisphosphate ( PIP2) to be hydrolyzed to inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DG). Therefore, the phosphatidylinositol (PLC) signalling pathway is also called the “dual messenger system”.

IP3 turns on the calcium channel and initiates signals downstream. Ca2+ binds to calmodulin (CaM) to form a Ca2+-CaM complex, which activates adenylate cyclase (AC) and phosphodiesterase (PDE); activates Ca2+-CaM dependent protein kinase. DAG activates PKC, phosphorylates serine/threonine residues of proteins, and produces different cellular responses, such as cell secretion, muscle contraction, cell proliferation and differentiation. The PLC-γ pathway is also involved in the differentiation of red blood cells.

3. cAMP/PKA signalling pathway

Cyclic adenosine monophosphate (cAMP) is an important intracellular signalling molecule, activates cAMP-dependent protein kinase A (PKA), and regulates cell differentiation. cAMP/PKA signalling can promote the adipogenic differentiation of MSCs and inhibit their osteogenic differentiation.

4. Via JAK-STAT

JAK is a tyrosine kinase whose main substrate is the STAT transcription factor. Activated STAT translocates to the nucleus and binds to the DNA sequence, thus regulating gene expression. The JAK-STAT pathway plays an important role in cell proliferation, apoptosis, and differentiation.

The main process of this signal pathway is as follows:

• The binding of the ligand to the receptor leads to receptor dimerization. Dimerized receptors activate JAK, JAK phosphorylation STAT. Phosphorylated STAT forms dimers that enter the nucleus and bind to DNA sequences to regulate gene expression.
• The JAK/STAT pathway plays an important role in the proliferation and differentiation of red blood cells.

5. PI3K-AKT-mTOR signalling pathway

The mammalian target of rapamycin (mTOR) is a conserved serine/threonine-protein kinase with two main forms: mTORC1 and mTORC2. Activated mTOR plays a key regulatory role in cell proliferation, differentiation, and metabolism. mTOR is primarily regulated by the PI3K/Akt/mTOR signalling pathway and the LKB1/AMPK/mTOR signalling pathway.

These two signalling pathways are the main pathways that regulate the proliferation and differentiation of testicular support cells. In the mTOR signalling pathway, deletion of the mTOR gene leads to a decrease in the number of testicular support cells. Studies have shown that PI3K-activated Akt kinase plays an important role in hematopoiesis. The PI3K pathway is also important in small intestinal stem cell regeneration and in promoting cell differentiation.

6. Wnt signalling pathway

The Wnt signalling pathway is highly conserved and the central component of this signalling pathway is β-catenin. When the Wnt signal is not activated, intracellular β-catenin is phosphorylated and degraded by the proteasome. When the Wnt protein binds to its receptor Frizzled and LRP, the β-catenin degradation complex is inactivated, β-catenin is released and accumulates in the cell. Accumulated β-catenin enters the nucleus and binds to transcription factor and T-cell factor/lymphocyte-enhancing factor (TCF/LEF) to initiate a series of proliferation-related genes.

6.1 Wnt signalling pathway and adipocyte differentiation

Activation of the Wnt signalling pathway can inhibit fat cell differentiation, and once this pathway is out of control, obesity can occur. The Wnt signalling pathway is activated by interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-α), which inhibits β-catenin degradation. β-catenin further inhibits downstream levels of C/EBP and PPAR, impairing or even preventing adipocyte differentiation.

Fat cell differentiation was also affected by the PPAR signalling pathway and the Hedgehog signalling pathway. The PPAR family plays an important role in fat differentiation and metabolism. Among them, PPARγ, as an important transcription factor for adipogenic differentiation, can accelerate adipocyte differentiation and deposition.

6.2 Wnt signalling pathway and cartilage differentiation

Activation of the Wnt signalling pathway can promote chondrocyte differentiation. During chondrogenic differentiation, the Wnt signalling pathway acts in conjunction with many other pathways. In cartilage differentiation, TGF-β and Wnt signalling pathways together promote cartilage differentiation from mesenchymal stem cells. In addition, the Wnt signalling pathway can promote small intestinal stem cell differentiation and cardiomyocyte differentiation, and promote the proliferation and differentiation of testicular support cells.

7. TGF-β Superfamily Signaling Pathway

The TGF-β superfamily regulates cell growth, proliferation, differentiation, migration, and apoptosis, regulates embryonic development, participates in the body’s immune response, and has multifunctional biological activities. The TGF-β signalling pathway affects the proliferation and differentiation of testicular support cells.

Summary

The interspersed repeat content of mammalian genomes has been best characterized in humans, mice, and cows. In this study, we carried out a de novo identification of repeated elements in the equine genome and identified previously unknown elements present at low copy numbers. The equine genome contains repeats typical of eutherian mammals but also has a significant number of hybrid repeats in addition to clade-specific long interspersed nuclear elements (LINEs).

The clade-specific LINE 1 (L1) repeats of Equus caballus Recombinant can be classified into approximately five subfamilies, three of which have undergone significant expansion. There are 1115 complete copies of this equine L1s, but of the 103 presumed active copies, 93 belong to a single subfamily, indicating a recent rapid expansion of this subfamily. We also analyzed both genome-wide simple sequence repeats (SSRs) and interspersed ones, finding that some repeat classes are spatially correlated with each other, as well as with G+C content and gene density.

On the basis of these spatial correlations, we have confirmed that recently described clade-specific versus ancestral genome territories can be defined by their repeat content. Correlations of clade-specific short interspersed nuclear elements were scattered throughout the genome and appear to have been extensively remodelled. In contrast, territories enriched by ancestral repetitions tended to be contiguous domains.

To determine whether these latter territories were evolutionarily conserved, we compared these results with a similar analysis of the human genome and observed enriched domains with similar ancestral repeats. These results indicate that evolutionarily conserved territories of the ancestral mammalian genome can be identified on the basis of repeat content alone. Interspersed repeats of different ages appear to be analogous to geological strata, allowing identification of ancient versus newly remodelled regions of mammalian genomes.

Purity: >85% (SDS-PAGE)

Target names: INS

Uniprot No.: P01310

Alternative Names: SIN; Insulin [Split into insulin B chain; insulin A chain]

Species: Equus caballus (Horse)

Expression Region: 1-30

Protein Length: Cytoplasmic Domain

Label information

The following labels are available.

• N-terminus His-tagged
• Without tags
• The type of label will be determined during the production process. If you have specified a tag type, let us know and we will develop the specified tag preferentially.

Form: Lyophilized powder

Buffer before lyophilization: Tris/PBS based buffer, 6% trehalose, pH 8.0

Reconstitution

We recommend that this vial be briefly centrifuged before opening to bring the contents to the bottom. Reconstitute protein in sterile deionized water at a concentration of 0.1-1.0 mg/mL. We recommend adding 5-50% glycerol (final concentration) and an aliquot for long-term storage at -20℃/-80℃. Our final default glycerol concentration is 50%. Customers could use it for reference.

Storage Conditions

Store at -20°C/-80°C upon receipt, need to be aliquoted for multiple uses. Avoid repeated cycles of freezing and thawing.

Shelf life

Shelf life is related to many factors, storage condition, buffer ingredients, storage temperature and the stability of the protein itself. Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C. The shelf life of the lyophilized form is 12 months at -20°C/-80°C.

Delivery time

The delivery time may differ depending on the way or location of purchase, consult your local distributors for the specific delivery time.

Note: All of our proteins are shipped with regular blue ice packs by default. If you request shipping with dry ice, please contact us in advance and additional fees will be charged.

Notes: Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.