Mouse Interleukin-3 Recombinant

HomeHematopoietins, IL-3 family, Recombinant Mouse Cytokines

Mouse Interleukin-3 Recombinant

$70.00 – $4,700.00

SKU: RKP01586 Categories: Hematopoietins, IL-3 family, Recombinant Mouse Cytokines Tags: Csfmu, il-3, IL3, interleukin-3, mcgf, P01586

Description

Methods (41)


accession	P01586

Source	Optimized DNA sequence encoding mouse Interleukin-3 mature chain was expressed in Escherichia Coli.
Molecular weight	Native mouse Interleukin-3 is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 15kDa. Recombinant mouse IL-3 is a monomer consisting of 135 amino acid residue subunits, and migrates as an approximately kDa protein under non-reducing conditions and reducing conditions in SDS-PAGE.
Purity	>98%, as determined by SDS-PAGE and HPLC
Biological Activity	The ED(50) was determined by the dose-dependent stimulation of the proliferation of mouse M-NFS-60 cells was found to be in the <0.05 ng/ml.
Protein Sequence	MVLASSTTSI HTMLLLLLML FHLGLQASIS GRDTHRLTRT LNCSSIVKEI IGKLPEPELK TDDEGPSLRN KSFRRVNLSK FVESQGEVDP EDRYVIKSNL QKLNCCLPTS ANDSALPGVF IRDLDDFRKK LRFYMVHLND LETVLTSRPP QPASGSVSPN RGTVEC
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-3 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.

Molecular function	Cytokine
Molecular function	Growth-factor

Methods


Generation of BMDC and BMMC For BMMC induction, 1×10⁶ BM cells were cultured supplemented with 5 ng/ml recombinant murine SCF and IL-3 for more than three weeks (>98% expressed c-kit and FcεRIα).
Mast cell culture from bone marrow Mouse bone marrow-derived cultured mast cells (BMCMC) were differentiated from femoral bone marrow by culture in medium supplemented with recombinant mouse IL-3 and stem cell factor(rmSCF, , ) as described ^−/− bone marrow showed similar granular morphology, levels of active tryptase, and expression of F_cεRIα and CD117, indicating that they mature similarly when cultured in the presence of IL-3 and SCF.
Effects of overexpression of Bmi1 on HSCs in vitro. Single CD34-LSK cells were sorted into 96-well microtiter plates containing the SF-O3 medium supplemented with 10% FBS and multiple cytokines (10 ng/ml SCF, 10 ng/ml TPO, 10 ng/ml IL-3, and 3 u/ml EPO) and allowed to form colonies.
Effects of overexpression of Bmi1 on HSCs in vitro. Single CD34-LSK cells were sorted into 96-well microtiter plates containing the SF-O3 medium supplemented with 10% FBS and multiple cytokines (10 ng/ml SCF, 10 ng/ml TPO, 10 ng/ml IL-3, and 3 u/ml EPO) and allowed to form colonies.
Generation and visualization of neutrophil extracellular traps (NETs) from myeloid cell lines and human primary neutrophils. EPRO cells were generated by differentiating the EML multipotent progenitor cell line with ATRA, GM-CSF and IL-3.
Induction of 2nd iPSCs All isolated somatic cells were cultured in the presence of Dox (2 ug/ml) for induction of 2^nd iPSCs. Fibroblasts and hematopoietic cells were cultured in ES/iPSC medium. FLCD45 were cultured in the presence of 10 ng/ml human TPO, 10 ng/ml mouse EPO, 10 ng/ml mouse IL-3, 10 ng/ml mouse IL-6, 10 ng/ml mouse Flt3 ligand, 10 ng/ml mouse GM-CSF, 10 ng/ml mouse VEGF and 50 ng/ml mouse SCF . HSCs, HPCs and MPs were cultured in the presence of 10 ng/ml human TPO, 10 ng/ml mouse IL-3, 10 ng/ml mouse IL-6 and 10 ng/ml mouse Flt3 ligand . Macrophages were cultured in the presence of 5 ng/ml M-CSF .
Induction of 2nd iPSCs All isolated somatic cells were cultured in the presence of Dox (2 ug/ml) for induction of 2^nd iPSCs. Fibroblasts and hematopoietic cells were cultured in ES/iPSC medium. FLCD45 were cultured in the presence of 10 ng/ml human TPO, 10 ng/ml mouse EPO, 10 ng/ml mouse IL-3, 10 ng/ml mouse IL-6, 10 ng/ml mouse Flt3 ligand, 10 ng/ml mouse GM-CSF, 10 ng/ml mouse VEGF and 50 ng/ml mouse SCF . HSCs, HPCs and MPs were cultured in the presence of 10 ng/ml human TPO, 10 ng/ml mouse IL-3, 10 ng/ml mouse IL-6 and 10 ng/ml mouse Flt3 ligand . Macrophages were cultured in the presence of 5 ng/ml M-CSF .
Biological activities of HoxB4 in Ba/F3 cells. 2×103 Ba/F3-HoxB4 cells were cultured with 5 ng/ml IL-3 (survival cytokine) in 96-well plates for 3 days and cell proliferation activity was assessed via MTT assay.
Lentiviral transduction of bone marrow cells Freshly isolated bone marrow (BM) cells were plated in Iscove's Modified Dulbecco's Medium (IMDM) (31980, , ) containing 5% fetal bovine serum (FBS), 1% penicillin/streptomycin, 200 mM glutamine, 1% non-essential amino acids, 1% sodium pyruvate, 50 μM 2-mercaptoethanol, stem cell factor (SCF, 10 ng/ml, , ), Flt3-L (10 ng/ml), IL-11 (10 ng/ml), thrombopoietin (TPO, 10 ng/ml), IL-6 (10 ng/ml), and IL-3 (10 ng/ml). Bone marrow cells were spin-infected with 8 μg/ml polyprene and lentivirus (MOI of 0.1–0.5), at 2,500 rpm for 90 min. BM cells were then incubated for 3 hours at 37°C, counted and injected into NOD.SCID mice.
Generation and isolation of MLL-AF9 leukemia cells MLL-AF9 mouse leukemias were generated in Dr. David Scadden's laboratory as described below. Actin-DsRED mice (JAX) were backcrossed for 10 generations onto C57BL/6J mice (JAX). These mice were sacrificed four days after injection with 150 mg/kg 5FU. BM cells were isolated from femurs and tibias, and red blood cells were lysed using ACK lysing buffer . Cells were incubated in RPMI supplemented with 20% FBS, 1% Pen-Strep, 6 ng/ml of IL-3 , 10 ng/ml of TPO , 10 ng/ml of IL-6 , at 37°C 5% CO2 overnight. MLL-AF9 was introduced by spin-infection (1 000 g for 90 minutes) using a retroviral vector (MSCV-MLL-AF9-neo) ⁶ live cells per mouse were injected into lethally irradiated (9Gy) C57BL/6 recipient mice 12 hours after viral infection. The mice were sacrificed once they became moribund. BM cells were isolated as described above, and subjected to ACK lysis. 200 000 cells were injected into sublethally irradiated (4.5 Gy)… MLL-AF9 mouse leukemias were generated in Dr. David Scadden's laboratory as described below. Actin-DsRED mice (JAX) were backcrossed for 10 generations onto C57BL/6J mice (JAX). These mice were sacrificed four days after injection with 150 mg/kg 5FU. BM cells were isolated from femurs and tibias, and red blood cells were lysed using ACK lysing buffer . Cells were incubated in RPMI supplemented with 20% FBS, 1% Pen-Strep, 6 ng/ml of IL-3 , 10 ng/ml of TPO , 10 ng/ml of IL-6 , at 37°C 5% CO2 overnight. MLL-AF9 was introduced by spin-infection (1 000 g for 90 minutes) using a retroviral vector (MSCV-MLL-AF9-neo) ⁶ live cells per mouse were injected into lethally irradiated (9Gy) C57BL/6 recipient mice 12 hours after viral infection. The mice were sacrificed once they became moribund. BM cells were isolated as described above, and subjected to ACK lysis. 200 000 cells were injected into sublethally irradiated (4.5 Gy) C57BL/6 recipients. Once these mice were moribund, ACK-lysed live BM cells were isolated as described above and 1×10⁶ cells/mouse were used for intramuscular injections. Read more