Human IL3 Baculovirus-Insect cells Overexpression Lysate

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Human IL3 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human IL3 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of IL3 protein (Cat: 11858-H08B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human IL3 (NP_000579.2) (Met 1-Phe 152) was expressed, fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human IL3 consists of 143 amino acids and predicts a molecular mass of 16.5 kDa. It migrates as an approximately 19 kDa band in SDS-PAGE under reducing conditions.

Human IL3 Baculovirus-Insect cells Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human IL3 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human IL-3 Overexpression Lysate; Human Interleukin-3 Overexpression Lysate; Human MCGF Overexpression Lysate; Human MULTI-CSF Overexpression Lysate

IL3 Background Information

IL3 (interleukin 3), also known as IL-3, is a potent growth promoting cytokine which belongs to the IL-3 family. IL3/IL-3 also belongs to the group of interleukins. Interleukins are produced by a wide variety of body cells. The function of the immune system depends in a large part on interleukins, and rare deficiencies of a number of them have been described, all featuring autoimmune diseases or immune deficiency. The majority of interleukins are synthesized by helper CD4+ T lymphocytes, as well as through monocytes, macrophages, and endothelial cells. They promote the development and differentiation of T, B, and hematopoietic cells. IL3/IL-3 is capable of supporting the proliferation of a broad range of hematopoietic cell types. It is involved in a variety of cell activities such as cell growth, differentiation and apoptosis. IL3/IL-3 has been shown to also possess neurotrophic activity, and it may be associated with neurologic disorders.
Full Name
interleukin 3
References
  • Meyer CG, et al.. (2011) IL3 variant on chromosomal region 5q31-33 and protection from recurrent malaria attacks. Hum Mol Genet. 20(6):1173-81.
  • Zambrano A, et al. (2010) IL-3 controls tau modifications and protects cortical neurons from neurodegeneration. Curr Alzheimer Res. 7(7):615-24.
  • Dentelli P, et al.. (2011) IL-3 is a novel target to interfere with tumor vasculature. Oncogene. 30(50):4930-40.
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