Active Prolactin (PRL)

LTH; Luteotropic Hormone

ACTIVITY TEST

PRL (prolactin), also known as luteotropin, is a hormone secreted from the pituitary gland and is best known for its role in enabling mammals to produce milk. PRL plays an essential role in metabolism, regulation of the immune system. According to reports, PRL also stimulates proliferation of certain cells, including MCF-7. Thus, proliferation assay of PRL was conducted using MCF-7 cells. Briefly, MCF-7 cells were seeded into triplicate wells of 96-well plates at a density of 2,000cells/well and allowed to attach overnight, then the medium was replaced with serum-free standard DMEM prior to the addition of various concentrations of PRL. After incubated for 72h, cells were observed by inverted microscope and cell
proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly,10µL of CCK-8 solution was added to each well of the plate, then the absorbance at 450nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37oC.
(A) MCF-7 cells cultured in DMEM, stimulated with 1ng/mL PRL for 72h;
(B) Unstimulated MCF-7 cells cultured in DMEM for 72h.
Figure. Cell proliferation of MCF-7 cells after stimulated with PRL.

The dose-effect curve of PRL was shown in Figure 2. It was obvious that PRL significantly promoted cell proliferation of MCF-7 cells. The ED50 for this effect is typically 3.709 to 8.973 ng/mL.
Figure. The dose-effect curve of PRL on MCF-7 cells.

PRL (prolactin), also known as luteotropin, is a hormone secreted from the pituitary gland and is best known for its role in enabling mammals to produce milk. PRL plays an essential role in metabolism, regulation of the immune system through activating its specific membrane-anchored receptor (PRLR). PRL can bind to PRLR, inducing JAK2 association that leads to downstream activation of multiple pathways that include STAT3, STAT5, PI3K, AKT, and ERK. A functional binding ELISA assay was conducted to detect the interaction of recombinant human PRL and recombinant rat JAK2. Briefly, PRL was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to JAK2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-PRL pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630 nm immediately. The binding activity of recombinant human PRL and recombinant rat JAK2 was shown in Figure 1, the EC50 for this effect is 4.5 ug/mL.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
The Journal of Biological Chemistry In Vivo Evidence for Epidermal Growth Factor Receptor (EGFR)-mediated Release of Prolactin from the Pituitary Gland Jcb: 39297
Journal of Clinical Neuroscience TLR9 expression is associated with prognosis in patients with glioblastoma multiforme ScienceDirect: S0967586811003572
Sensors and Actuators B: Chemical Determination of prolactin hormone in serum and urine using an electrochemical immunosensor based on poly(pyrrolepropionic acid)/carbon nanotubes hybrid modified electrodes Sciencedirect:S0925400514000720
Veterinary Word Effect of extended photoperiod during winter on growth and onset of puberty in Murrah buffalo heifers Pubmed:27051212
Journal of Dairy Science Effect of thermal stress on physiological, hormonal and haematological parameters in Tharparkar and Karan Fries calves publication:306322503
Stem Cell Research & Therapy Differentiation of human umbilical cord Wharton’s jelly-derived mesenchymal stem cells into endometrial cells pubmed:29096715
Stem Cell Research & Therapy The role of mesenchymal stem cells in chemotherapy-induced gonadotoxicity Pubmed:30021657
Frontiers in Molecular Neuroscience Impact of Triclosan on Female Reproduction through Reducing Thyroid Hormones to Suppress Hypothalamic Kisspeptin Neurons in Mice Pubmed:29403355
Toxicology and Applied Pharmacology The antipsychotics sulpiride induces fatty liver in rats via phosphorylation of insulin receptor substrate-1 at Serine 307-mediated adipose tissue insulin resistance Pubmed:29551354
Journal of Cellular Biochemistry The prolactin‐release inhibitor paeoniflorin suppresses proliferation and induces apoptosis in prolactinoma cells via the mitochondria‐dependent pathway Pubmed:29388711
Journal of Cellular and Molecular Medicine Metformin inhibits growth and prolactin secretion of pituitary prolactinoma cells and xenografts Pubmed: 30334324
BMC Veterinary Research Hormonal and metabolic indicators before and after farrowing in sows affected with postpartum dysgalactia syndrome Pubmed: 30404636
International Journal of Basic and Clinical Endocrinology Expression of prolactin receptors in the duodenum, kidneys and skeletal system during physiological and sulpiride-induced hyperprolactinaemia Pubmed: 30143940
MORPHOLOGY AND PATHOMORPHOLOGY Morphological and Biochemical Characteristics of Prostate Hyperplasia during Sulpiride Treatment Pubmed: 32152847
JOURNAL OF DAIRY SCIENCE Effect of heat stress during the early and late dry period on mammary gland development of Holstein dairy cattle Pubmed: 32684470
BioMed Research International Pathophysiological Changes in Female Rats with Estrous Cycle Disorder Induced by Long-Term Heat Stress Pubmed: 32685488
Vet Microbiol Prolactin affects the disappearance of ALV-J viremia in vivo and inhibits viral infection 34391195
Antioxidants (Basel) Effects of a Nanoencapsulated Moringa Leaf Ethanolic Extract on the Physiology, Metabolism and Reproductive Performance of Rabbit Does during Summer 34439574
Nutrients Yogurt Enriched with Inulin Ameliorated Reproductive Functions and Regulated Gut Microbiota in Dehydroepiandrosterone-Induced Polycystic Ovary Syndrome Mice Pubmed:35057459
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