Human IL-13 HEK293 Overexpression Lysate: Product Information
This Human IL-13 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IL-13 protein (Cat: 10369-H01H) from the overexpression lysate was verified.
A DNA sequence encoding the mature form of human IL13 (AAK53823.1) (Gly 21-Asn 132) was fused with the Fc region of human IgG1 at the N-terminus.
The recombinant human Fc/IL13 chimera is a disulfide-linked homodimeric protein. The reduced monomer consists of 349 amino acids and predicts a molecular mass of 39 kDa. The apparent molecular mass of rhIL13/Fc monomer is approximately 50-55 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Human IL-13 HEK293 Overexpression Lysate: Usage Guide
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.
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.
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human IL-13 HEK293 Overexpression Lysate: Synonyms
Human ALRH Overexpression Lysate; Human IL-13 Overexpression Lysate; Human P600 Overexpression Lysate
IL-13 Background Information
Interleukin 13 (IL-13) is a single-chain glycosylated polypeptide, which belongs to the IL-13/IL-4 family. IL-13 protein is secreted by many cell types, but especially by T helper type 2 (Th2) cells. IL-13 exerts its effects through a multi-subunit receptor comprising the alpha chain of the IL-4 receptor (IL-4Rα) and at least one of two known IL-13-specific binding chains (IL-13 Rα1 and IL-13 Rα2). As a cytokine, IL-13 protein is critical in regulating inflammatory, immune responses, and diseases. Also, it inhibits the production of pro-inflammatory cytokines and chemokines, and thus down-regulates macrophage activity. IL-13 protein and antibody are more importantly implicated as a central mediator of immunoregulatory processes in various cell types.
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