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|Baculovirus-Insect Cell lysate that Human XCL2 transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).|
|A DNA sequence encoding the human XCL2 (Q9UBD3) (Met 1-Gly 114) was fused with a polyhistidine tag at the C-terminus.|
|The secreted recombinant human XCL2 comprises 103 amino acids and has a predicted molecular mass of 11.7 kDa. The apparent molecular mass of rh XCL2 is approximately 18-20 kDa in SDS-PAGE under reducing conditions.|
|Cell lysate was prepared by homogenization 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.|
|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.|
|12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.|
|Samples are stable for up to twelve months from date of receipt.|
|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. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.|
|1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).|
|Store at 4℃. After re-dissolution, aliquot and store at -80℃.|
|Western blot (WB): Use at an assay dependent dilution.|
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
XCL2 is a small cytokine belonging to the XC chemokine family. Chemokines are a group of cytokines. Members of this group of so-called C-Chemokines belong to the SCY family of cytokines and are designated XCL (L for ligand) followed by a number. XCL2 is the new designation of SCM-1-beta. XCL2 is highly related to another chemokine called XCL1. XCL2 gene is located on chromosome 1 in humans. XCL2 is predominantly expressed in activated T cells, but can also be found at low levels in unstimulated cells. XCL2 induces chemotaxis of cells expressing the chemokine receptor XCR1. XCL2 gene has been proposed to participate in pathways (Chemokine signaling pathway, Cytokine-cytokine receptor interaction) and processes (blood circulation, signal transduction, chemotaxis, immune response). XCL2 are expected to have molecular function (chemokine activity) and to localize in various compartments (extracellular space, extracellular region). A putative protein interactor has been described (XCR1). The spliced mRNA putatively encodes a good protein, containing small cytokines (intecrine / chemokine), interleukin-8 like domain; the complete protein appears to be secreted.