- ApplicationsConjugation, Puri
Peanut Lectin validated data
Application Information: Peanut Lectin
|Full Name||Peanut agglutinin lectin, Arachis hypogaea |
|Synonyms||PNA lectin, PNA, Arachis hypogaea, Peanut Lectin, Peanut glutinin ctin, Arachis hypogaea, Peanut agglutinin, Arachis hypogaea lectin, Peanut agglutinin lectin|
|Product Description||Arachis hypogaea lectin against Gal beta 1-3 GalNac|
|Specificity||Gal beta 1-3 GalNac|
|Background||Peanut lectin (PNA) is an identical tetrameric carbohydrate free protein with MW of 110 kDa. Thomsen-Friedenreich antigen, T-antigen (Galβ1, 3GalNAc) is present on blood group M &N glycoproteins (after removal of sialic acid with neuraminidase), glyconjugates (Mucin type), gangliosides and many glycolipids. T antigen is rarely expressed on normal coloncytes whereas cells of malignant, premalignant cells express this antigen. Peanut lectin has widely been used to detect T antigen in malignant and premalignant cells. Peanut lectin contains one atom of Ca++ and Mg++ per unit. |
Inhibiting/Eluting sugars: 200 mM GalactoseCarbohydrate-Binding Specificity of PNA: Galβ1, 3 GalNAc>>>>>GalNAc>Gal
|Target||Gal beta 1-3 GalNac|
|Application Note||Conjugation to Sepharose 4B (solid phase columns) to purify mucin type glycoproteins. For conjugation azide should be removed. Dilute in buffer containing 0.1 mM calcium chloride. The optimum dilution should be determined by the individual lab.|
|Storage Buffer||10 mM bicarbonate, 150 mM NaCl, pH 8.2, 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.