Human Interleukin 1 beta Recombinant
Categories: Interleukin-1 familyRecombinant Human Cytokines$70.00 – $3,500.00
Description
Accession
P01584
Source
Optimized DNA sequence encoding Human Interleukin-1 beta mature chain was expressed in Escherichia Coli.
Molecular weight
Mature Human IL-1 beta, is generated by the proteolytic removal of the signal peptide and propeptide.The molecule has a calculated molecular mass of approximately17 kDa. Recombinant IL-1 beta is a monomer protein consisting of 154 amino acid residue subunits. Recombinant IL-1 beta migrates as an approximately 17 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity
>97%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) as determined by thedose-dependent proliferation of murine D10S cells, was found to be ≤0.1 pg/ml, corresponding to a specific activity of ≥1 x107units/mg.
Protein Sequence
MAEVPELASE MMAYYSGNED DLFFEADGPK QMKCSFQDLD LCPLDGGIQL RISDHHYSKG FRQAASVVVA MDKLRKMLVP CPQTFQENDL STFFPFIFEE EPIFFDTWDN EAYVHDAPVR SLNCTLRDSQ QKSLVMSGPY ELKALHLQGQ DMEQQVVFSM SFVQGEESND KIPVALGLKE KNLYLSCVLK DDKPTLQLES VDPKNYPKKK MEKRFVFNKI EINNKLEFES AQFPNWYIST SQAENMPVFL GGTKGGQDIT DFTMQFVSS
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation
IL-1 beta was lyophilized from a 0.2 μm filtered PBS pH7.0.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.
Storage
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 at2° -8° 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.
Usage
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Interactor
P29466
Biological Process
Molecular function
Molecular function
Molecular function
Methods
Nampt and NMN increase NAD level and ameliorate NAD depletion.
- Human islets and INS-1E cells were stimulated with NMN [100 µM] or Nampt [2.5 ng/ml] for a short time (2 or 3 h) or a long time exposure (48 h).
In vitro Suspension Culture
- After informed consent had been obtained, cord blood from healthy volunteers was collected into sterile heparinized tubes.
- Mononuclear cells were obtained by centrifugation on Lymphoprep density gradient.
- Ex vivo cultures were set up with cord blood MNCs at 3×106 cells/mL in medium'>StemSpan medium supplemented with 50 ng/mL thrombopoietin (TPO) (Sunshine Pharmaceutical Company Limited), 20 ng/mL interleukin-3 (IL-3) , 50 ng/mL stem cell factor (SCF) and 50 ng/ml IL-6 using 75 cm2 tissue culture flasks (Corning).
- At indicated time points, the cultured cells were harvested, washed, and prepared for immediate infusion.
Chondrocyte cell culture studies
- Cell culture experiments were conducted using, and C28/I2 immortalized human chondrocytes, and involved first plating cells in 6-well culture plates at 200,000 viable cells per well in 1:1 DMEM/F-12 media supplemented with 10% FBS.
- After 48 hours, the culture medium was replaced with 2.0 ml of serum-free 1:1 MEM/F-12 and incubated for 5 hours prior to treatment with recombinant human (rh) MATN3 protein (100 ng/ml or 200 ng/ml) and/or rhIL-1β (5.0 ng/ml) (ocky , , ) for 8 to 36 hours, unless otherwise stated.
- To optimize the induction of catabolic proteases (that is, MMP-13, ADAMTS family) by IL-1 in both primary and immortalized chondrocytes, we used serum-free media for the duration of the treatment period as in a previous study [
Th1-induced vascular endothelial growth factor but not angiopoietin-like-4 expression is dependent on hypoxia-inducible factor-1.
- Changes in mRNA in response to IL-1β or TNFα of (a) vascular endothelial growth factor and (b) angiopoietin-like -4 expressed as fold-change relative to levels in the siLuc transfected untreated controls set as 1.0 (dotted line).
Neutrophil adhesion to IL-1β-activated EC or PC monolayers.
- Freshly isolated (naïve) neutrophils were seeded onto EC or PC monolayers previously activated with IL-1β for 4 or 24 hr in Sykes-Moore chambers and allowed to adhere prior to counting.
Cell Culture
- VSMCs, EL4, HEK, and HeLa cells were cultured in DMEM and Jurkat and THP-1 cells in RPMI 1640, all supplemented with penicillin, streptomycin, L-glutamine, and 10% FCS.
- Human monocyte-derived macrophages were differentiated as described previously (2, clarified, and stored at −80°C.
- Cells were also made necrotic by incubation with 7-BIO (25 μM) or digitonin (0.1%) (data not shown) or by overnight hypoxic exposure.
- To activate inflammasomes, cells were treated with LPS (1 μg/ml; 4 hr), followed by ATP (5 mM) or Nigericin (20 μM) for 30 min.
- Calpain activity was determined with Calpain-Glo .
- VSMCs, HEK, and THP-1 cells were transfected with pcDNA3 with nucleofection or FugeneHD .
PB1-F2 modulates NF-κB signaling.
- Forty eight hours post-transfection, the cells were either induced or not induced with 25 µg ml−1 Poly , 20 ng ml−1 TNFα, 1 µg ml−1 LPS or 10 ng ml−1 IL-1β for five hours, cell extracts were made and the luciferase and galactosidase activites were measured.
IL-1.
- Relative expression of miR-125b in normal (n = 3) and osteoarthritic (n = 7) chondrocytes stimulated with IL-1β for 24 hours.
IL-1.
- Relative expression of miR-125b in normal (n = 3) and osteoarthritic (n = 7) chondrocytes stimulated with IL-1β for 24 hours.
Upregulation of pro-IL-1ß and conversion to mature IL-1ß following exposure to A2E.
- Membranes were probed with anti-human IL-1ß, which detects both the pro and mature form of the cytokine.
