Description
Accession
P16581
Source
Optimized DNA sequence encoding Human E-Selectin (CD62E/ELAM1) extracellular domain[22-556] was expressed in CHO cells.
Molecular weight
Recombinant human Selectin\CD62E, generated by the proteolytic removal of the signal peptide and propeptide, is a monomer protein consisting of 534 amino acid residue subunits. The molecule has a calculated molecular mass of approximately 59kDa. E-Selectin migrates due to glycosylation as an approximately kDa protein under non-reducing conditions and as a 85 kDa protein under reducing conditions in SDS-PAGE.
Purity
>95%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determined by the dose-dependent stimulation of the proliferation of monkeyMBr-5 cells was found to be in the range of.0-40.0 ng/ml.
Protein Sequence
MIASQFLSAL TLVLLIKESG AWSYNTSTEA MTYDEASAYC QQRYTHLVAI QNKEEIEYLN SILSYSPSYY WIGIRKVNNV WVWVGTQKPL TEEAKNWAPG EPNNRQKDED CVEIYIKREK DVGMWNDERC SKKKLALCYT AACTNTSCSG HGECVETINN YTCKCDPGFS GLKCEQIVNC TALESPEHGS LVCSHPLGNF SYNSSCSISC DRGYLPSSME TMQCMSSGEW SAPIPACNVV ECDAVTNPAN GFVECFQNPG SFPWNTTCTF DCEEGFELMG AQSLQCTSSG NWDNEKPTCK AVTCRAVRQP QNGSVRCSHS PAGEFTFKSS CNFTCEEGFM LQGPAQVECT TQGQWTQQIP VCEAFQCTAL SNPERGYMNC LPSASGSFRY GSSCEFSCEQ GFVLKGSKRL QCGPTGEWDN EKPTCEAVRC DAVHQPPKGL VRCAHSPIGE FTYKSSCAFS CEEGFELHGS TQLECTSQGQ WTEEVPSCQV VKCSSLAVPG KINMSCSGEP VFGTVCKFAC PEGWTLNGSA ARTCGATGHW SGLLPTCEAP TESNIPLVAG LSAAGLSLLT LAPFLLWLRK CLRKAKKFVP ASSCQSLESD GSYQKPSYIL
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation
Interleukin-6 was lyophilized from a 0.2 μm filtered solution in.5% glycine,.5% sucrose,.01% Tween80, mM Glutamic acid, pH.5.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers
Storage
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at2° -8° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Interactor
O00592
Interactor
Interactor
Interactor
Biological Process
Methods
Selectin-mediated slow leukocyte rolling is abolished in patients after CPB.
- The rolling velocity of neutrophils in whole blood from healthy control volunteers (dashed line), on-pump patients (straight line) and OPCAB patients (dotted line) was measured after induction of anesthesia , following administration of protamine , and 24 hours after the end of surgery using microflow chambers coated with E-selectin or P-selectin alone and in combination with ICAM-1.
Selectin-mediated slow leukocyte rolling is abolished in patients after CPB.
- The rolling velocity of neutrophils in whole blood from healthy control volunteers (dashed line), on-pump patients (straight line) and OPCAB patients (dotted line) was measured after induction of anesthesia , following administration of protamine , and 24 hours after the end of surgery using microflow chambers coated with E-selectin or P-selectin alone and in combination with ICAM-1.
CPB during cardiac surgery increases chemokine-induced leukocyte arrest.
- Flow chambers coated with P-selectin/ICAM-1 or P-selectin/ICAM-1/Interleukin-8 were perfused with whole blood from healthy control volunteers (black bars) or from cardiac surgery patients operated with the use of CPB (on-pump patients, grey bars) or from patients operated in OPCAB technique (OPCAB patients, white bars) after induction of anesthesia , after the administration of protamine , and 24 hours after the end of the surgery .
Solid phase binding assay
- Maxisorp 96-well plates (U96 from VWR, , The ) were coated overnight with purified Tsp-C18 carrying chains of variable lengths (chain chain and Core-Tsp1-C18) diluted 1∶10 with phosphate buffered saline (PBS), pH 7.5, as well as with the serum-free CM collected from 293-EBNA cells expressing the Short-ColXVIII.
- After washing with PBS- 0.05% Tween 20, the wells were blocked with 5% skimmed milk powder in PBS for 1 h. After washing, a dilution range of recombinant L-selectin-IgM chimera 2SO4.
- Absorbance was measured at 450 nm with a microplate reader.
- All incubations were done at room temperature in a volume of 100 µl/well.
