Human SIGLEC5 HEK293 Overexpression Lysate: Product Information
This Human SIGLEC5 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of SIGLEC5 protein (Cat: 11798-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human SIGLEC5 (O15389) (Met1-Thr 434) was expressed, fused with the Fc region of human IgG1 at the C-terminus.
The recombinant human SIGLEC5 /Fc is a disulfide-linked homodimer. The reduced monomer comprises 659 amino acids and has a predicted molecular mass of 73.4 kDa. The apparent molecular mass of the protein is approximately 74-93 kDa in SDS-PAGE under reducing conditions.
Human SIGLEC5 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 SIGLEC5 HEK293 Overexpression Lysate: Alternative Names
Human CD170 Overexpression Lysate; Human CD33L2 Overexpression Lysate; Human OB-BP2 Overexpression Lysate; Human OBBP Overexpression Lysate; Human OBBP2 Overexpression Lysate; Human SIGLEC-5 Overexpression Lysate; Human SIGLEC5 Overexpression Lysate
SIGLEC5 Background Information
SIGLEC5 contains 2 Ig-like C2-type (immunoglobulin-like) domains and 1 Ig-like V-type (immunoglobulin-like) domain. It belongs to the immunoglobulin superfamily and SIGLEC (sialic acid binding Ig-like lectin) family. SIGLEC5 is expressed by monocytic/myeloid lineage cells. It is found at high levels in peripheral blood leukocytes, spleen, bone marrow and at lower levels in lymph node, lung, appendix, placenta, pancreas and thymus. It is also expressed by monocytes and neutrophils but absent from leukemic cell lines representing early stages of myelomonocytic differentiation. SIGLEC5 is a putative adhesion molecule that mediates sialic-acid dependent binding to cells. It binds equally to alpha-2,3-linked and alpha-2,6-linked sialic acid. The sialic acid recognition site may be masked by cis interactions with sialic acids on the same cell surface.
sialic acid binding Ig like lectin 5
Angata T, et al. (2006) Discovery of Siglec-14, a novel sialic acid receptor undergoing concerted evolution with Siglec-5 in primates. FASEB J. 20(12):1964-73.
Avril T, et al. (2005) Siglec-5 (CD170) can mediate inhibitory signaling in the absence of immunoreceptor tyrosine-based inhibitory motif phosphorylation. J Biol Chem. 280(20):19843-51.
Erickson-Miller CL, et al. (2003) Characterization of Siglec-5 (CD170) expression and functional activity of anti-Siglec-5 antibodies on human phagocytes. Exp Hematol. 31(5):382-8.
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