| Description : |
Raf-1 is the cellular homolog of viral Raf gene (v-Raf). The encoded protein is a MAP Kinase kinase kinase (MAP3K), which functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated, the cellular RAF1 protein can phosphorylate to activate the dual specificity protein kinases MEK1 and MEK2, which in turn phosphorylate to activate the serine/threonine specific protein kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in this gene are associated with Noonan syndrome 5 and LEOPARD syndrome 2. |
| Source : |
Sf21 |
| Species : |
Human |
| Tag : |
GST |
| Protein length : |
306-end |
| Form : |
0.564mg/ml of enzyme in 50mM Tris/HCl, pH7.5, 150mM NaCl, 0.1mM EGTA, 0.03% Brij-35, 270mM sucrose, 1mM benzamidine, 0.2mM PMSF, 0.1% 2-mercaptoethanol. Frozen solution. |
| Bio-activity : |
113212U/mg, where one unit of Raf-1 activity = 1 unit of MAPK2 activity which in turn is defined as 1nmol phosphate incorporated into 0.33mg/ml MBP per minute at 30 centigrade with a final ATP concentration of 100μM. Note the activity is determined using a triple linked assay which involves the activation of MEK1 by RAF-1, followed by the subsequent activation of MAPK2 by the activated MEK1. |
| Molecular Mass : |
65 kDa |
| AA Sequence : |
1 MSPILGYWKI KGLVQPTRLL LEYLEEKYEE HLYERDEGDK WRNKKFELGL EFPNLPYYID 61 GDVKLTQSMA IIRYIADKHN MLGGCPKERA EISMLEGAVL DIRYGVSRIA YSKDFETLKV 121 DFLSKLPEML KMFKDRLCHK TYLNGDHVTH PDFMLYDALD VVLYMDPMCL DAFPKLVCFK 181 KRIEAIPQID KYLKSSKYIA WPLQGWQATF GGGDHPPKSD LVPRGSQPKT PVPAQRERAP 241 VSGTQEKNKI RPRGQRDSSD DWEIEASEVM LSTRIGSGSF GTVYKGKWHG DVAVKILKVV 301 DPTPEQFQAF RNEVAVLRKT RHVNILLFMG YMTKDNLAIV TQWCEGSSLY KHLHVQETKF 361 QMFQLIDIAR QTAQGMDYLH AKNIIHRDMK SNNIFLHEGL TVKIGDFGLA TVKSRWSGSQ 421 QVEQPTGSVL WMAPEVIRMQ DNNPFSFQSD VYSYGIVLYE LMTGELPYSH INNRDQIIFM 481 VGRGYASPDL SKLYKNCPKA MKRLVADCVK KVKEERPLFP QILSSIELLQ HSLPKINRSA 541 SEPSLHRAAH TEDINACTLT TSPRLPVF |
| Purity : |
Purity 89.8% by SDS-PAGE and Coomassie blue staining. |
| Applications : |
Drug Discovery & Development |
| Storage : |
6 months at -70 centigrade. |
| Handling Advice : |
Rapidly thaw the vial under cold water and immediately place on ice. Aliquot unused material into prechilled micro-centrifuge tubes and immediately snap-freeze the vials in liquid nitrogen prior to re-storage at -70 centigrade. |
| Use/Stability : |
On receipt of material store at -70 centigrade. Unopened reagent is stable for a minimum of 6 months from date of shipment when stored at recommended storage temperature. Avoid repeat freeze/thaw cycles. For maximum recovery of product, centrifuge original vial prior to removing the cap. |
