This Mouse CD99L2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD99L2 protein (Cat: 50526-M02H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse CD99L2 (NP_612182.1) extracellular domain (Met 1-Ala 164) was fused with the Fc region of human IgG1 at the C-terminus.
The secreted recombinant mouse CD99L2/Fc is a disulfide-linked homodimeric protein. The reduced monomer comprises 380 amino acids and has a predicted molecular mass of 42 kDa. As a result of glycosylation, the apparent molecular mass of rmCD99L2/Fc monomer is approximately 55-65 kDa in SDS-PAGE under reducing conditions.
Mouse CD99L2 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.
CD99 antigen-like protein 2, also known as MIC2-like protein 1, CD99L2 and MIC2L1, is a single-pass type I membrane protein which belongs to theCD99 family. CD99L2 is expressed in brain, heart, lung, liver, spleen, kidney, stomach, small intestine, skeletal muscle, ovary, thymus, testis and uterus. Lower expression of CD99L2 is seen in thymus. It is also expressed in E18 uterus and placenta. CD99 and CD99L2 were required for leukocyte extravasation in the cremaster after stimulation with tumor necrosis factor-alpha, where the need for PECAM-1 is known to be bypassed. CD99 and CD99L2 act independently of PECAM-1 in leukocyte extravasation and cooperate in an independent way to help neutrophils overcome the endothelial basement membrane. CD99L2 may function as a homophilic adhesion molecule. It functions in leukocyte-endothelial cell interactions during leukocyte extravasation, and in particular, at the diapedesis step. CD99L2 does not seem to be involved in docking of leukocytes to the vessel wall or in lymphocyte diapedesis.
CD99 molecule-like 2
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