UEA I Lectin (Biotin)
- ApplicationsELISA, IHC, WB
UEA I Lectin (Biotin) validated data
UEA I Lectin (Biotin)
|Full Name||Ulex europaeus Lectin|
|Synonyms||UAE, Ulex europaeus, UAE I Lectin, Fucose lectin, UAE I, Ulex europaeus lectin, UAE Lectin|
|Product Description||Ulex europaeus lectin against fucose blood group H (Biotin)|
|Specificity||Fucose blood group H|
|Background||The anti-H (O) hemagglutinating activity of Ulex europaeus has been used widely to confirm blood group O activity. UEA I binds to many glycoproteins and glycolipids containing α –linked fucose residue. The purified lectin appears to be a dimer of two distinct polypeptide chains associated by noncovalent forces. This lectin does not react with Lea active blood group substance. The native protein has pI=6.0-6.1 and exhibits a Molecular weight 60,000-68,000 during gel filtration on Sephadex column. This lectin will hemagglutinate fresh blood group O cells at 2-5μg/ml.Inhibiting/Eluting sugars: 50-100 mM L-fucoseCarbohydrate-Binding Specificity of UEA I: L-Fuα1,2Galβ1,4GlcNAcβ1,6R|
|Target||Fucose blood group H|
|Applications||ELISA, IHC, WB|
|Application Note||Histochemistry 1: 100-1:1000; WB 1:200-1:2,000; ELISA 1:500-1:2,500. Dilute biotinylated lectin in PBS containing 0.1 mM calcium chloride ions. For Histochemistry the tissues are processed same as for Immunohistochemistry, after blocking step biotinylated lectin is applied followed by streptavidin conjugated to enzyme and chromogen. For WB incubate membrane with blocking protein followed by biotinylated lectin, streptavidin enzyme conjugate and chromogen. For ELISA the target proteins are absorbed on ELISA plate at a concentration of 5-10 µg/ml (50-100 µl), followed by blocking with protein solution, Biotinylated lectin, Streptavidin enzyme conjugate, ELISA substrate. Please refer to Histochemistry, WB and ELISA protocol for detail information.The optimum dilution should be determined by the individual lab.|
|Conjugation Note||Affinity purified UEA I is conjugated with Biotin and purified in gel filtration column|
|Storage Buffer||10 mM phosphate, 150 mM NaCl, pH 7.6, 0.1 mM Calcium chloride and 0.05% sodium azide|
|Storage Instruction||Store at 2-8°C.|
|Notes||For in vitro research use only. Not intended for any diagnostic or therapeutic purpose.|
Why is the observed Western Blot band size different from predicted size?
The predicted M.W. is based on protein sequence analysis; however, some factors might lead to an observed band size that is different from the predicted size. The reasons might include:
1.Post-translational modification (PTM):
a. Some post-translational modifications might lead to increased protein size, including
phosphorylation, acetylation, methylation, glycosylation, sumoylation, ubiquitination,
b. Some post-translational modifications might lead to decreased protein size including
phosphatidylethanolamine conjunction (e.g. LC3-II)
c. Some proteins may be cleaved to form an active or mature form; this process will
lead to a decreased protein size (e.g. Notch activation, Caspase activation, etc.)
d. Some websites provide useful PTM information
iv.CBS data sets http://www.cbs.dtu.dk/databases/
v.CBS prediction Servers http://www.cbs.dtu.dk/services/
2.mRNA splice variants (Isoforms):
Through alternative splicing, one gene can generate different proteins with different M.W. Regulation of alternative splicing depends upon cell type, conditions, etc.
Some proteins could form dimers or multimers, increasing the M.W. This phenomenon usually can be found in reducing gel condition; however, strong interactions may still be seen with higher molecular weight proteins even in denaturing gel.
The observed size could also potentially be influenced by the protein charge
Different species likely have different protein sequence and PTM, which can lead to a different protein M.W.
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