ProDots Recombinant Human M-CSF Protein Summary
Pre-aliquoted, lyophilized protein dots for use in cell culture mediumFeatures and Benefits
- Rolls out of bottle for easy cell culture medium preparation
- Eliminates time spent aliquoting
- One protein dot can be used to make 500 mL of 10 ng/mL cell culture medium
ProDots® proteins are R&D Systems industry-leading quality, highly bioactive proteins packaged into easy-to-use lyophilized “dots”. Designed to be stable at 4 ºC and to dissolve instantly, ProDots® proteins can be rolled into cell culture media to quickly produce media with a desired working protein concentration.
ProDots® proteins are designed to reduce cell culture media preparation time while ensuring optimal protein recovery and bioactivity. Unlike standard lyophilized protein, which is shipped as a diffuse residue within a vial, ProDots® proteins are neatly bundled into the shape of a ball. In this form ProDots® proteins can be rolled directly into cell culture media. ProDots® proteins rehydrate instantly in media to maximize the recovery of purchased protein and to reliably reconstitute it at a desired concentration for use in cell culture media. ProDots® proteins can be conveniently stored at room temperature where they will remain stable for 6 months or at 4 ºC where they will remain stable for 12 months. The accessible storage and high stability of ProDots® proteins makes them ideal for quick and easy preparation of cell culture media.
Glu33 - Ser190, with an N-terminal Met
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Reconstitution||For a stock solution, reconstitute at 50-500 μg/mL in sterile PBS, or simply roll ProDot® directly into cell culture medium.
|Shipping||The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
• 6 months from date of receipt at room temperature.
• 12 months from date of receipt at 2-8 °C as supplied.
• 1 month at 2-8 °C under sterile conditions after reconstitution.
• 3 months at -20 to -80 °C under sterile conditions after reconstitution.
ProDots®Recombinant Human M-CSF stimulates cell proliferation of the M-NFS-60 mouse myelogenous luekemia lymphoblast cell line in a dose-dependent manner. The ED50 for this effect is 0.5-1.5 ng/mL.
1 μg/lane of ProDots®Recombinant Human M-CSF was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized with silver staining, showing bands at 19 kDa and 35 kDa, respectively.
M-CSF, also known as CSF-1, is a four-alpha-helical-bundle cytokine that is the primary regulator of macrophage survival, proliferation and differentiation. M-CSF is also essential for the survival and proliferation of osteoclast progenitors. M-CSF also primes and enhances macrophage killing of tumor cells and microorganisms, regulates the release of cytokines and other inflammatory modulators from macrophages, and stimulates pinocytosis. M-CSF increases during pregnancy to support implantation and growth of the decidua and placenta. Sources of M-CSF include fibroblasts, activated macrophages, endometrial secretory epithelium, bone marrow stromal cells and activated endothelial cells. The M-CSF receptor (c-fms) transduces its pleotropic effects and mediates its endocytosis. M-CSF mRNAs of various sizes occur. Full length human M-CSF transcripts encode a 522 amino acid (aa) type I transmembrane (TM) protein with a 464 aa extracellular region, a 21 aa TM domain, and a 37 aa cytoplasmic tail that forms a 140 kDa covalent dimer. Differential processing produces two proteolytically cleaved, secreted dimers. One is an N- and O- glycosylated 86 kDa dimer, while the other is modified by both glycosylation and chondroitin-sulfate proteoglycan (PG) to generate a 200 kDa subunit. Although PG-modified M-CSF can circulate, it may be immobilized by attachment to type V collagen. Shorter transcripts encode M-CSF that lack cleavage and PG sites and produce an N-glycosylated 68 kDa TM dimer and a slowly produced 44 kDa secreted dimer. Although forms may vary in activity and half-life, all contain the N-terminal 150 aa portion that is necessary and sufficient for interaction with the M-CSF receptor. The first 223 aa of mature human M-CSF shares 88%, 86%, 81% and 74% aa identity with corresponding regions of dog, cow, mouse and rat M-CSF, respectively. Human M-CSF is active in the mouse, but mouse M-CSF is reported to be species-specific.
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