分泌型卷曲相关蛋白1(SFRP1)活性蛋白
[ PROPERTIES ]
Source: Prokaryotic expression. Host: E. coli
Residues: Val8~Asp153
Tags: N-terminal His-tag
Purity: >95%
Endotoxin Level: <1.0EU per 1μg (determined by the LAL method). Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 0.05% sarcosyl
and 5% trehalose. Applications: Cell culture; Activity Assays. (May be suitable for use in other assays to be determined by the end user.)
Predicted isoelectric point: 9.1
Predicted Molecular Mass: 17.9kDa
Accurate Molecular Mass: 18kDa as determined by SDS-PAGE reducing conditions.
[ USAGE ]
Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0mg/mL. Do not vortex.
[ STORAGE AND STABILITY ]
Storage: Avoid repeated freeze/thaw cycles.
Store at 2-8oC for one month. Aliquot and store at -80oC for 12 months.
Stability Test: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37oC for 48h, and no obvious degradation and precipitation wereobserved. The loss rate is less than 5% within the expiration date underappropriate storage condition.
[ SEQUENCE ]
[ ACTIVITY ]
Secreted frizzled-related protein 1, also known as SFRP1 is a member of theSFRP family that contains a cysteine-rich domain homologous to the putativeWnt-binding site of Frizzled proteins. It acts as a biphasic modulator of Wntsignaling, counteracting Wnt-induced effects at high concentrations and promotingthem at lower concentrations. As a tumor suppressor, SFRP1 expression markedlyinhibited tumor cell growth in culture, soft agar and xenografts in athymic nudemice. Besides, Wingless Type MMTV Integration Site Family, Member 2 (WNT2)has been identified as an interactor of SFRP1, thus a binding ELISA assay wasconducted to detect the interaction of recombinant rat SFRP1 and recombinant ratWNT2. Briefly, SFRP1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to WNT2-coated microtiter wellsand incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1hwith anti-SFRP1 pAb, then aspirated and washed 3 times. After incubation withHRP labelled secondary antibody, wells were aspirated and washed 3 times. Withthe addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. Thebinding activity of SFRP1 and WNT2 was shown in Figure 1, and this effect was ina dose dependent manner.
To measuered the ability of SFRP1 on tumor suppression, liver cancer HepG2cells were seeded into triplicate wells of 96-well plates at a density of 5,000cells/well and allowed to attach, replaced with serum-free overnight, then themedium was replaced with 2% serum standard DMEM prior to the addition ofvarious concentrations of recombinant rat SFRP1. After incubated for 96h, cellswere observed by inverted microscope and cell proliferation was measured by CellCounting Kit-8 (CCK-8). Briefly, 10µL of CCK-8 solution was added to each well ofthe plate, then the absorbance at 450nm was measured using a microplate readerafter incubating the plate for 1-4 hours at 37℃. Suppression of HepG2 cells afterincubation with SFRP1 for 96h observed by inverted microscope was shown inFigure 2. Cell viability was assessed by CCK-8 assay after incubation withrecombinant SFRP1 for 96h. The result was shown in Figure 3. It was obvious thatSFRP1 significantly decreased cell viability of HepG2 cells.
[ IDENTIFICATION ]
[ IMPORTANT NOTE ]
The kit is designed for research use only, we will not be responsible for any issue if the kit was used in clinical diagnostic or any other procedures.
特别提示:本公司的所有产品仅可用于科研实验,严禁用于临床医疗及其他非科研用途!以实际收货产品说明书为准,网站说明书仅供参考。