Erythrina cristagalli Lectin (ECA, ECL) (Agarose)
Cat# E3453-19A-2ml
Size : 2ml
Brand : US Biological
Conjugate
AgaroseGrade
Molecular Biology GradeApplications
EShipping Temp
Blue IceStorage Temp
4°C Do Not FreezeErythrina cristagalli lectin is a 54,000D glycoprotein consisting of two different subunits of approximately 28,000 and 26,000D. It has a specificity toward galactose residues and appears to have the highest binding activity toward galactosyl (b -1,4) N-acetylglucosamine.
This lectin has been reported to be useful for the isolation of human natural killer (NK) cells using a negative selection panning technique. Human NK cells appear to lack accessible surface carbohydrate structures required for binding ECL and, unlike other mononuclear cells, do not adhere to ECL-coated culture dishes. Since this procedure involves a negative selection panning technique, a high recovery of viable NK cells can be obtained. The adherent cells can also be recovered by incubation in galactose or lactose. The carbohydrate structure to which ECL binds is frequently found in membrane and serum glycoproteins of mammalian origin. Sialic acid substitution on this structure appears to prevent the lectin from binding. This specificity offers an opportunity to utilize agarose bound ECL to isolate or fractionate mammalian glycoproteins. ECL has also been reported to be mitogenic to human peripheral T lymphocytes but is not mitogenic toward human peripheral B lymphocytes, mouse splenocytes or mouse thymocytes. Agarose bound Erythrina Cristagalli Lectin is prepared from affinity-purified lectin. Heat stable, cross-linked 4% agarose beads with a molecular weight exclusion limit of about 2x10e7 are used as the solid-phase matrix to which the lectin is covalently bound. The attachment of the lectin to the solid phase is carefully controlled in order to preserve the activity of the lectin as well as to minimize conformational changes of the bound lectin which might result in nonspecific ionic or hydrophobic interactions. The technique developed to couple lectins to agarose provides a very hydrophilic spacer arm between the protein and the matrix. This ensures maximum expression of the carbohydrate binding activity of the lectin. The linkage is very stable over a range of pH values and, unlike cyanogen bromide linkages, proteins are not leached off the gel by Tris or other routinely used buffers. In addition, residual charges generated during cyanogen bromide conjugation which can produce nonspecific binding are not present on the gel following our coupling procedure.
This lectin has been reported to be useful for the isolation of human natural killer (NK) cells using a negative selection panning technique. Human NK cells appear to lack accessible surface carbohydrate structures required for binding ECL and, unlike other mononuclear cells, do not adhere to ECL-coated culture dishes. Since this procedure involves a negative selection panning technique, a high recovery of viable NK cells can be obtained. The adherent cells can also be recovered by incubation in galactose or lactose. The carbohydrate structure to which ECL binds is frequently found in membrane and serum glycoproteins of mammalian origin. Sialic acid substitution on this structure appears to prevent the lectin from binding. This specificity offers an opportunity to utilize agarose bound ECL to isolate or fractionate mammalian glycoproteins. ECL has also been reported to be mitogenic to human peripheral T lymphocytes but is not mitogenic toward human peripheral B lymphocytes, mouse splenocytes or mouse thymocytes. Agarose bound Erythrina Cristagalli Lectin is prepared from affinity-purified lectin. Heat stable, cross-linked 4% agarose beads with a molecular weight exclusion limit of about 2x10e7 are used as the solid-phase matrix to which the lectin is covalently bound. The attachment of the lectin to the solid phase is carefully controlled in order to preserve the activity of the lectin as well as to minimize conformational changes of the bound lectin which might result in nonspecific ionic or hydrophobic interactions. The technique developed to couple lectins to agarose provides a very hydrophilic spacer arm between the protein and the matrix. This ensures maximum expression of the carbohydrate binding activity of the lectin. The linkage is very stable over a range of pH values and, unlike cyanogen bromide linkages, proteins are not leached off the gel by Tris or other routinely used buffers. In addition, residual charges generated during cyanogen bromide conjugation which can produce nonspecific binding are not present on the gel following our coupling procedure.
Applications:
Suitable for use in ELISA. Other applications not tested.
Recommended Dilution:
Optimal dilutions to be determined by the researcher.
Binding Capacity:
≥2.3mg asialo-fetuin/ml of gel
Inhibiting/Eluting Sugar:
200mM lactose 3mg lectin/ml gel
Activity:
2x10e7
Storage and Stability:
May be stored at 4°C. For long-term storage, aliquot and store at 4°C. Do not freeze. Aliquots are stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.
Source
Erythrina cristagalli seeds
Concentration
~1mg/ml
Form
Supplied as a liquid in 10mM HEPES, pH 7.5, 0.15M sodium chloride, 0.1mM Ca++, 20mM lactose, 0.08% sodium azide. Labeled with Agarose.
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.