Human STIM1 HEK293 Overexpression Lysate: Product Information
This Human STIM1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of STIM1 protein (Cat: 11434-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human STIM1 (NP_003147.2) extracellular domain (Met 1-Asp 213) was expressed, with a polyhistidine tag at the C-terminus.
The recombinant human STIM1 consists of 202 amino acids and predictes a molecular mass of 23.3 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh STIM1 is approximately 33-38 kDa due to glycosylation.
Human STIM1 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 STIM1 HEK293 Overexpression Lysate: Synonyms
Human D11S4896E Overexpression Lysate; Human GOK Overexpression Lysate; Human IMD10 Overexpression Lysate; Human STRMK Overexpression Lysate; Human TAM Overexpression Lysate; Human TAM1 Overexpression Lysate
STIM1 Background Information
Stromal interaction molecule 1, also known as STIM1 and GOK, is a cell membrane, a single-pass type I membrane protein and a endoplasmic reticulum membrane protein. STIM1 / GOK is ubiquitously expressed in various human primary cells and tumor cell lines. It contains one EF-hand domain and one SAM (sterile alpha motif) domain. STIM1 / GOK plays a role in mediating Ca2+ influx following depletion of intracellular Ca2+ stores. It acts as Ca2+ sensor in the endoplasmic reticulum via its EF-hand domain. Upon Ca2+ depletion, STIM1 / GOK translocates from the endoplasmic reticulum to the plasma membrane where it activates the Ca2+ release-activated Ca2+ (CRAC) channel subunit, TMEM142A / ORAI1. Transfection of STIM1 / GOK into cells derived from a rhabdoid tumor and from a rhabdomyosarcoma that do not express detectable levels of STIM1 can induce cell death, suggesting a possible role in the control of rhabdomyosarcomas and rhabdoid tumors. Defects in STIM1 are the cause of immune dysfunction with T-cell inactivation due to calcium entry defect type 2 (IDTICED2) which is an immune disorder characterized by recurrent infections, impaired T-cell activation and proliferative response, decreased T-cell production of cytokines, lymphadenopathy, and normal lymphocytes counts and serum immunoglobulin levels.
stromal interaction molecule 1
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Manji S.S. et al., 2000, Biochim. Biophys. Acta 1481: 147-55.
Williams R.T. et al., 2002, Biochim. Biophys. Acta. 1596: 131-7.
Spassova M.A. et al., 2006, Proc. Natl. Acad. Sci. USA. 103: 4040-5.
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