Optimized DNA sequence encoding HumanProlactin mature chain was expressed in Escherichia Coli.
Human Prolactin is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately23 kDa. Recombinant Prolcatin is a monomeric protein consisting of200 amino acid residue subunits, and migrates as an approximately23 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
>98%, as determined by SDS-PAGE and HPLC
The ED(50) was determined by the dose-dependent stimulation of the proliferation of monkeyMBr-5 cells was found to be in the range of.0-40.0 ng/ml.
MNIKGSPWKG SLLLLLVSNL LLCQSVAPLP ICPGGAARCQ VTLRDLFDRA VVLSHYIHNL SSEMFSEFDK RYTHGRGFIT KAINSCHTSS LATPEDKEQA QQMNQKDFLS LIVSILRSWN EPLYHLVTEV RGMQEAPEAI LSKAVEIEEQ TKRLLEGMEL IVSQVHPETK ENEIYPVWSG LPSLQMADEE SRLSAYYNLL HCLRRDSHKI DNYLKLLKCR IIHNNNC
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Recombinant human Prolactin was lyophilized from a.2 μm filtered NaHCO3 solution.
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
In Vitro Differentiation
- To examine the ability of hBSCs to spontaneously differentiate, breastmilk cells were initially grown as spheroids (see above).
- By day 4–7, some cells had attached.
- The remaining spheroids were transferred into new wells where adherent cells appeared in 1–2 days.
- Both the initial and subsequent attached cells were cultured for another 2–3 weeks, with media changes every 3–5 days.
- For directed differentiation, primary and first- to third-passage breastmilk cell cultures were incubated in differentiation media at 37°C and 5% CO2 for 3–4 weeks.
- For mammary differentiation, cells were incubated in Roswell Park medium'>Memorial medium'>Institute medium (RPMI) 1640 with
l-glutamine supplemented with 20% FBS, 4 μg/ml insulin , 20 ng/ml epidermal growth factor (EGF) , 0.5 μg/ml hydrocortisone , 5% antibiotic-antimycotic, and 2 μL/ml fungizone.
- For osteoblastic and adipogenic differentiation, cells were incubated in NH OsteoDiff or NH Adipodiff medium, respectively .
- For chondrocyte differentiation,…