TIGIT lysate

Human TIGIT HEK293 Overexpression Lysate

• Applications: Western Blot
• Synonyms: TIGIT; Human TIGIT HEK293 Overexpression Lysate; Human TIGIT HEK293 Cell Lysate (WB positive control); Human VSIG9 Overexpression Lysate; Human VSTM3 Overexpression Lysate; Human WUCAM Overexpression Lysate; T cell immunoreceptor with Ig and ITIM domains; TIGIT lysate

• Host: HEK293 Cells
• Form/Format: Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.; 1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).

• Applicable Applications for TIGIT lysate: Western Blot (WB)
• Species: Human
• Sequence Information: A DNA sequence encoding the human TIGIT (NP_776160.2)(Met1-Pro141) was expressed with the Fc region of mouse IgG2a at the C-terminus.
• Preparation Method: Cell lysate was prepared by homogenization 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 (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
• 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 200uL pure water and boil for 2-5 min.; 3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
• Preparation and Storage: Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.

Related Product Information for TIGIT lysate

Description: This Human TIGIT overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TIGIT protein from the overexpression lysate was verified.

Background: TIGIT, also known as V-set and transmembrane domain-containing protein 3 (VSTM3) or V-set and immunoglobulin domain-containing protein 9 (VSIG9) is a new surface protein containing an immunoglobulin variable domain, a transmembrane domain and an immunoreceptor tyrosine-based inhibitory motif (ITIM). TIGIT is expressed on regulatory, memory, activated T cells and NK cells. It binds PVR with high affinity, and PVRL2 with lower affinity, but not PVRL3. Knockdown of TIGIT with siRNA in human memory T cells did not affect T cell responses, however, TIGIT inhibits NK cytotoxicity directly through its ITIM. TIGIT suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. The binding of PVR to TIGIT on human dendritic cells enhanced the production of IL-1 and diminished the production of IL-12p4. Also, TIGIT counter inhibits the NK-mediated killing of tumor cells and protects normal cells from NK-mediated cytotoxicity thus providing an "alternative self" mechanism for MHC class I inhibition.

Product Categories/Family for TIGIT lysate

ITIM/ITAM Immunoreceptors and Related Molecules Cancer Drug Targets

References

Ota T., et al.,(2004), Complete sequencing and characterization of 21,243 full-length human cDNAs. Nat. Genet. 36:40-45.Bechtel S., et al., (2007), The full-ORF clone resource of the German cDNA consortium.BMC Genomics 8:399-399.Yu X., Harden K., et al.,(2009), The surface protein TIGIT suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Nat. Immunol. 10:48-57.