- ApplicationsConjugation, Puri
ConA Lectin validated data
Application Information: ConA Lectin
|Full Name||Concanavalin A, Canavalia ensiformis|
|Synonyms||Concanavalin A, Concanavalin A, Canavalia ensiformis, Canavalia ensiformis lectin, ConA lectin, Canavalia ensiformis, ConA, Concanavalin A lectin, Con A lectin|
|Product Description||Canavalia ensiformis lectin against alpha-linked mannose and glucose|
|Specificity||alpha-linked mannose and glucose|
|Background||Concanavalin A is a lectin protein (MW 104kDa), homotetramer 26 kDa; originally extracted from the jack-bean, Canavalia ensiformis. It binds specifically to certain structures found in various sugars α-mannosyl and α-glucosyl residues in glycoproteins. It was the first lectin to be available on a commercial basis and is widely used in biology and biochemistry to characterize glycoproteins and other sugar-containing entities. It is also used to purify macromolecules in lectin affinity chromatography. Concanavalin A interacts with diverse receptors containing mannose carbohydrates (serum and membrane glycoproteins).ConA agglutinate strongly erythrocytes without being blood group specific. Normal cell re agglutinated after trypsinisation. ConA is a also a lymphocyte mitogen.ConA reacts with many bacteria, like E. coli Dictyostelium discoideum et B. substilis It is also widely believed to be involved in the interaction between alpha-mannosyl oligosaccharides on the surface of the HIV virus and the human T cell lymphocyte. Inhibiting/Eluting sugars: 200 α-mM α-methyl mannoside / 200 mM α-methyl glucoside mixture.|
Carbohydrate-Binding Specificity of Con A: (Manα1,2Manα1,2Man > Manα1,2Man > α-Man > α-Glc > αGlcNAc
|Target||alpha-linked mannose and glucose|
|Application Note||Conjugation to Sepharose 4B (solid phase columns) to purify glycoproteins, and viral antigen isolation. 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, 0.1 mM Calcium chloride, 0.01mM manganese chloride and 0.05% sodium azide. (Con A has an Isoelectric point of about pH 4.5-5.5 and requires calcium or manganese ions at each of its four saccharide binding sites; THESE|
|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.