Application Note
UDP Glucose Ceramide Glucosyltransferase (UGCG) is an enzyme which catalyzes the first glycosylation step in glycosphingolipid biosynthesis. It belongs to the glycosyltransferase 2 family. UGCG is widely expressed and transcription is upregulated during keratinocyte differentiation. Besides, Large Multifunctional Peptidase 2 (LMP2) has been identified as an interactor of UGCG, thus a binding ELISA assay was conducted to detect the interaction of recombinant human UGCG and recombinant human LMP2. Briefly, UGCG were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to LMP2-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-UGCG pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of UGCG and LMP2 was in a dose dependent manner.
Observed MW
16 kDa.
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Lys39~Leu171 (NP_003349.1)
Expression System
E. coli
Purity
> 95%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Synonyms
UDP-glucose ceramide glucosyltransferase , GCS , GLCT1
Background
This gene encodes an enzyme that catalyzes the first glycosylation step in the biosynthesis of glycosphingolipids, which are membrane components containing lipid and sugar moieties. The product of this reaction is glucosylceramide, which is the core structure of many glycosphingolipids. [provided by RefSeq, Dec 2014]
Database
Research Area