Mouse NBL1 HEK293 Overexpression Lysate: Product Information
This Mouse NBL1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NBL1 protein (Cat: 50976-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse NBL1 (Q61477) (Met 1-Asp 178) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant mouse NBL1 comprises 173 amino acids and has a calculated molecular mass of 18.8 kDa. As a result of glycosylation, the apparent molecular mass of the recombinant protein is approximately 28 kDa in SDS-PAGE under reducing conditions.
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.
Mouse NBL1 HEK293 Overexpression Lysate: Alternative Names
Mouse D4H1S1733E Overexpression Lysate; Mouse DAN Overexpression Lysate; Mouse Dana Overexpression Lysate; Mouse NO3 Overexpression Lysate
NBL1 Background Information
The Dan (Differential screening-selected gene aberrative in neuroblastoma, also known as N3) gene was first identified as the putative rat tumor suppressor gene and encodes a protein structurally related to Cerberus and Gremlin in vertebrates. It is a founding member of the DAN family of secreted proteins, acts as an inhibitor of cell cycle progression and is closely involved in retinoic acid-induced neuroblastoma differentiation. There are at least five mammalian protein members in the evolutionarily conserved Dan family including DAN, Gremlin/DRM, Cer1 (Cerberus-related), Dante and PRDC (protein related to DAN and cereberus), and share the C-terminal cystine-knot motif. As a secreted glycoprotein, DAN is a member of a class of glycoproteins shown to be secreted inhibitors of the transforming growth factor-beta (TGF-beta) and bone morphogenic protein pathways. It binds to BMPs and preventing their interactions with signaling receptor complexes, and accordingly regulates the processes of embryonic development and tissue differentiation. DAN gene product may have an important role in regulation of the entry of cells into the S phase. In addition, DAN gene product possesses an ability to revert phenotypes of transformed rat fibroblasts and represents a candidate tumour suppressor gene for neuroblastoma.
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