This Human I-309 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of I-309 protein (Cat: 10057-H01H) from the overexpression lysate was verified.
A DNA sequence encoding the mature form of human CCL1 (NP_002972.1) (Lys 24-Lys 96) was fused with the Fc region of human IgG1 at the N-terminus.
The recombinant human Fc/CCL1 is a disulfide-linked homodimeric protein. The reduced monomer consists of 310 amino acids and predicts a molecularmass of 35 kDa. As a result of glycosylation, the rhCCL1/Fc monomer migrates as an approximately 38-42 kDa band in SDS-PAGE under reducing conditions.
Human I-309 HEK293 Overexpression Lysate: 使用指南
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).
稳定性 & 储存条件
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 I-309 HEK293 Overexpression Lysate: 别称
Human I-309 Overexpression Lysate; Human P500 Overexpression Lysate; Human SCYA1 Overexpression Lysate; Human SISe Overexpression Lysate; Human TCA3 Overexpression Lysate
CCL1 or chemokine (C-C motif) ligand 1, also known as I-39 or TCA-3, is a member of the chemokine (C-C motif) ligand family. The C-C kemokines have two cysteines nearby the amino terminus. There have been at least 27 distinct members of this subgroup reported for mammals, called C-C chemokine ligands (CCL)-1 to 28. I-39/CCL1/TCA-3 interacts with the G protein-linked transmembrane chemokine receptors CCR8, and induces biochemical events that may result in the control of chemotaxis, proliferation, apoptosis and adhesion. It has been demonstrated that I-39/CCL1/TCA-3 displays chemotactic activity for monocytes and other cell types such as NK cells and dendritic cells, but not for neutrophils. Furthermore, as the only known physiological ligand for CCR8, I-39/CCL1/TCA-3 was identified as a potent inhibitor of HIV-1 envelope-mediated cell-cell fusion and virus infection. I-39/CCL1/TCA-3 induce significant levels of LTC4 from elicited eosinophils.
chemokine (C-C motif) ligand 1
Miller MD, et al. (1992) The human cytokine I-309 is a monocyte chemoattractant. Proc Natl Acad Sci. 89 (7): 2950-4.
Roos RS, et al. (1997) Identification of CCR8, the receptor for the human CC chemokine I-309. J Biol Chem. 272 (28): 17251-4.
Oliveira SH, et al. (2002) Increased responsiveness of murine eosinophils to MIP-1beta (CCL4) and TCA-3 (CCL1) is mediated by their specific receptors, CCR5 and CCR8. J Leukoc Biol. 71(6): 1019-25.