Human ULBP1 HEK293 Overexpression Lysate

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Human ULBP1 HEK293 Overexpression Lysate: Product Information

Product Description
This Human ULBP1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ULBP1 protein (Cat: 10679-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the mature form of human ULBP1 (NP_079494.1) (Met 1-Gly 216) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The mature form of recombinant human ULBP1 comprises 202 amino acids and predicts a molecular mass of 23.8 kDa. The apparent molecular mass of rhULBP1 is approximately 28-32 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Human ULBP1 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.

Human ULBP1 HEK293 Overexpression Lysate: Alternative Names

Human RAET1I Overexpression Lysate

ULBP1 Background Information

UL16-binding proteins (ULBP) or retinoic acid early transcripts-1 (RAET1) are ligands to the activating receptor, NKG2D. Ten members of the human ULBP/RAET1 gene family have been identified to encode for potentially functional proteins, and have tissue-specific expressions. ULBP1, also known as RAET1I and NKG2DL1, together with at least ULBP 2 and 3, are well-known ligands for NKG2D, and activate multiple signaling pathways in primary NK cells, resulting in the production of cytokines and chemokines. ULBP1 is expressed in T-cells, B-cells, erythroleukemia cell lines and in a wide range of tissues including heart, brain, lung, liver and bone marrow, as well as some tumor cells. As an unconventional member of the MHC class I family, ULBP1 function in immune responses, especially in cancer and infectious diseases. Unlike other ULBP members, ULBP1 is able to interact with soluble CMV glycoprotein UL16 in CMV infected cells. The interaction with UL16 blocked the interaction with the NKG2D receptor, and thus might escape the immune surveillance. Furthermore, UL16 also causes ULBP1 to be retained in the ER and cis-Golgi apparatus so that it does not reach the cell surface. The ULBP1 regulation may have implications for development of new therapeutic strategies against cancer cells.
Full Name
UL16 binding protein 1
References

    1.  Rölle, A. et al., 2003, J Immunol. 171(2): 902-908.  

    2.  López-Soto, A. et al., 2006, J Biol Chem. 281(41): 30419-30430.      

    3.  Song, H. et al., 2006, Cell Immunol. 239(1): 22-30.  

    4.  Eisele, G. et al., 2006, Brain. 129 (9): 2416-2425.   

    5.  Romphruk, AV. et al., 2009, Immunogenetics. 61(9): 611-617. 

    6.  Sutherland, C.L. et. al., 2002, J. Immunol. 168: 671-679.

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