Cynomolgus CD200RLa HEK293 Overexpression Lysate

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Cynomolgus CD200RLa HEK293 Overexpression Lysate: Product Information

Product Description
This Cynomolgus CD200RLa overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD200RLa protein (Cat: 11576-C02H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Cynomolgus
Sequence Information
A DNA sequence encoding the Cynomolgus (Macaca fascicularis) CD200RLa extracellular domain (Met 1-Leu 240) was as fused with the Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant cynomolgus CD200RLa/Fc is a disulfide-linked homodimer. The reduced monomer consists of 460 amino acids and has a calculated molecular mass of 51.3 kDa. The apparent molecular mass of the CD200RLa/Fc monomer is approximately 70-85 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Cynomolgus CD200RLa HEK293 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.

Cynomolgus CD200RLa HEK293 Overexpression Lysate: Alternative Names

Cynomolgus CD200RLa Overexpression Lysate

CD200RLa Background Information

Cell surface glycoprotein CD2 receptor 2, also known as Cell surface glycoprotein CD2 receptor 1-like, Cell surface glycoprotein OX2 receptor 2, CD2 receptor-like 2, CD2Rla, CD2R1L and CD2R2, is a single-pass type I membrane protein which belongs to theCD2R family. CD2R1L / CD2R2. It contains oneIg-like C2-type (immunoglobulin-like) domain and oneIg-like V-type (immunoglobulin-like) domain. CD2 is a transmembrane protein delivering immunoregulatory signals after engagement of CD2R. A family of CD2Rs exist ( CD2R1, CD2R2, CD2R3, CD2R4 ) with different tissue expression and functional activity. In the presence of anti-CD2R2 / CD2R3 monoclonal antibodies (mAbs), bone-marrow cells cultured in the presence of (interleukin [IL]-4+granulocyte-macrophage colony-stimulating factor) differentiate into dendritic cells (DCs), which induce CD4+CD25+ Treg. Interaction between the relatively ubiquitously expressed molecule CD2 and one of its receptors, CD2R1, resulted in direct suppression of alloreactivity, engagement of alternate receptors led instead to altered differentiation of dendritic cells (DCs) from marrow precursors, which could in turn foster development of Foxp3(+) regulatory T cells. Unlike anti-CD2R1, anti-CD2R2 both promotes development of DCs with capacity to induce Treg and directly augments thymocyte production of Treg.
Full Name
CD200 receptor 1 like
References
  • Wright G.J., et al.,(2003), Characterization of the CD200 receptor family in mice and humans and their interactions with CD200. J. Immunol. 171:3034-3046.
  • Gorczynski R.M., et al., (2004), Structural and functional heterogeneity in the CD200R family of immunoregulatory molecules and their expression at the feto-maternal interface.Am. J. Reprod. Immunol. 52:147-163.
  • Muzny D.M., et al.,(2006), The DNA sequence, annotation and analysis of human chromosome 3.Nature 440:1194-1198.
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