Mouse Interleukin-1 alpha Recombinant

///Mouse Interleukin-1 alpha Recombinant

Mouse Interleukin-1 alpha Recombinant

$70.00$4,700.00


accession P01582


Source Optimized DNA sequence encoding Mouse Interleukin-1 alpha mature chain was expressed in Escherichia Coli.
Molecular weight Mature Mouse IL-1 alpha, is generated by the proteolytic removal of the signal peptide and propeptide. The molecule has a calculated molecular mass of approximately kDa. Recombinant IL-1a is a monomer protein consisting of amino acid residue subunits and migrates as an approximately18 kDa protein under non-reducing conditions and reducing conditions in SDS-PAGE.
Purity >95%, as determined by SDS-PAGE and HPLC
Biological Activity The ED(50) was determined byacell proliferation assay using the murine helper T cell line, D10.G4., was foundto be10.0 pg/ml.

Protein Sequence MAKVPDLFED LKNCYSENED YSSAIDHLSL NQKSFYDASY GSLHETCTDQ FVSLRTSETS KMSNFTFKES RVTVSATSSN GKILKKRRLS FSETFTEDDL QSITHDLEET IQPRSAPYTY QSDLRYKLMK LVRQKFVMND SLNQTIYQDV DKHYLSTTWL NDLQQEVKFD MYAYSSGGDD SKYPVTLKIS DSQLFVSAQG EDQPVLLKEL PETPKLITGS ETDLIFFWKS INSKNYFTSA AYPELFIATK EQSRVHLARG LPSMTDFQIS
Endotoxin Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation Recombinant mouse Interleukin-1 alpha was lyophilized from a.2 μm filtered PBS solution.
Reconstitution 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.
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 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.
Usage This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.


Biological Process Inflammatory-response
Molecular function Cytokine
Molecular function Mitogen
Molecular function Pyrogen

Methods

Materials

  • Tetramethylammonium hydroxide was from Shanghai Lingfeng Chemical Reagent Co Ltd, Shanghai, China.
  • Other materials were RPMI 1640 medium, penicillin, and streptomycin , fetal bovine serum (, , , the ), recombinant murine granulocyte-macrophage colony-stimulating factor, recombinant murine interleukin-4, CD11c, B7-1 (CD80), B7-2(CD86), MHC II, and chemokine receptor 7 (eBioscience, San Diego, CA), tumor necrosis factor-α, interleukin-1β, and interleukin-6 , prostaglandin E2 , a Prussian blue staining kit .
  • Annexin V and propidium iodide , and the Cell Counting Kit-8 .

Early p38-induced IL-1β stimulates the later expression of the antiinflammatory cytokines IL-10 and TGFβ in cultured macrophages.

30 ng/ml recombinant IL-1β
  • As in B, cells were treated with an IL-1β neutralizing antibody, control IgG antibody, or recombinant IL-1β for 24 h (in this case, in the absence of LPS).

Experimental agents

  • Tetramethylammonium hydroxide was obtained from Lingfeng Chemical Reagent Co Ltd .
  • medium'>RPMI medium 1640, penicillin and streptomycin were from .
  • Fetal bovine serum (, , , the ), recombinant murine granulocyte-macrophage colony-stimulating factor, recombinant murine interleukin-4, tumor necrosis factor-α (TNF-α), interleukin-1β, interleukin-6 , prostaglandin E2 were also used, as well as a Prussian blue staining kit and isoflurane .
  • Triton-X , and normal goat serum, rabbit anti-GFP probes, and goat antirabbit Alexa 488 nm were also used.

IL-2/IL-21 and upregulation of IL-21R expression replace CD4+ T cell help of CD8+ T cell expansion in vitro.

IL-21R
  • IL-21R expression on CD8+ T cells stimulated with aAPC/mOKT3 in the presence or absence of CD4+ T cells was studied by flow cytometry.

Less proinflammatory cytokine induction was evident in IL-1 KO mice after SCI.

IL-1β
  • IL-1β levels in the spinal cords of wild-type mice (open squares and solid line) were drastically increased from the 1st dpo and sustained through to the 14th dpo.

Mammalian cell inflammatory cytokine levels during exposure to test bacteria.

Recombinant IL-8
  • Exposure supernatants were filtered and used in liquid array bead assays for IL-8 or IL-1β, IL-6 and TNF-α .

T-cell polarization.

  • Splenic cells (2 × 106/mL) from age 4 to 6 weeks naïve NOD.BDC2.5.FoxP3GFP.DTR mice were stimulated with p79 peptide (0.5 μmol/L) under Th1 or Th17 conditions for 4 days.
  • For polarization into Th1 cells, the stimulation was carried out in the presence of recombinant (r)IL-12 (10 ng/mL) and anti–IL-4 (10 μg/mL [11B11]).
  • For Th17 polarization, the culture was supplemented with recombinant transforming growth factor-β (3 ng/mL), rIL-6 (20 ng/mL), anti–IFN-γ (10 μg/mL [4–6A4]), and anti–IL-4 (10 μg/mL [11B11]) antibodies and rIL-23 (20 ng/mL& ) (only for the last 2 days).

Localization and activity of SpyCEP.

recombinant IL-8
  • SDS-PAGE and silver staining were performed after digestion of IL-8 (10 µg/ml) in the presence of 10 µg/ml of chloramphenicol.

Cell culture and infection

  • BEAS-2B or MH-S cells grown to 90% confluence were infected with RV or UV-RV at multiplicity of infection (MOI) of 1 or equal volume of sham (media from uninfected HeLa cells) and incubated for 90 min at 33°C.
  • Infection media was replaced with fresh media and the incubation continued for another 22 h. Cells were then infected with NTHi at MOI of 10 or treated with media alone, centrifuged at 500×g for 5 min and incubated at 37°C for 3 h. Media was collected for determination of IL-8 (for BEAS-2B cells) or KC, MIP-2 and TNF-α (for MH-S cells).
  • In some experiments, cells were infected with RV in the presence of 5 µM lactacystin or 10 to 100 ng/ml IL-1 receptor antagonist .
  • Primary cells were infected apically with RV, UV-RV or sham at 1 MOI

Cell lines and cell culture

  • 293T ( .
  • number CRL-11268) and NIH3T3 ( .
  • number CRL-1658) cells were both cultured in medium'>Dulbecco's medium'>modified medium'>Eagle's medium (DMEM) supplemented with 10% fetal calf serum (FCS) and 1% Penicillin-Streptomycin with 5% CO2 at 37°C.
  • Bone marrow cells (BM) isolated from wild type (wt) C57BL/6 mice were cultured in StemSpan Serum-free expansion medium supplemented with 1% glutamine, 1% penicillin-streptomycin, 100 ng/mL each murine stem cell factor , human Flt3-ligand and human interleukin 11 , and 10 ng/mL murine interleukin 3 .
  • The KSHV-positive PEL cell line BCBL-1