Erythrina Cristagalli Lectin (ECL)

Cat. No.: E1452704

GRAU & PUREZA BioReagent, Native

Visão geral

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.

This lectin has been reported to be useful for the isolation of human natural killer (NK) cells using a negative selection panning technique (protocol is provided below). 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.

Erythrina cristagalli lectin consists of two different subunits of approximately 28 kDa and 26 kDa.

Inhibiting/Eluting Sugar: 200 mM lactose

Specifications

Molecular Weight	54 kDa
Extinction Coefficient	1.3
Inhibiting or Eluting Sugar	Lactose
Storage Instructions	2–8 °C
Sugar Specificity	Terminal type 2 LacNAc, and terminal type 2 LacdiNAc
Usage Summary	Although many buffers can be employed for reconstituting this lectin, we recommend 10 mM HEPES buffered saline, 0.1 mM CaCl₂, pH 8.5. For preserving solutions stored at 4 ºC, 0.08% sodium azide can be used.
Applications	Glycobiology, Mitogenic Stimulation
Conjugate	Unconjugated

User Guide: Enrichment of NK Cells by Panning on Erythrina Cristagalli Plates

This procedure is a modification of the method for NK cell isolation described in the following reference: Koren HS, et al. 1985. Novel Approaches for Enrichment of NK Cells. Macrophage Biology. A variety of methods are used to obtain enriched fractions of NK cells, such as column adherence techniques, Percoll discontinuous gradients, and monoclonal antibodies. The following procedure utilizes the lectin isolated from Erythrina Cristagalli Lectin, taking advantage of its ability to discriminate between NK cells and other mononuclear cells in a negative selection technique. 

Procedure: 

1. Bacteriological plastic plates (60 mm) are coated with 2 ml of Erythrina Cristagalli lectin in PBS, pH 7.8, at a concentration of 250 μg/ml for 1 h at 24°C. 

2. Decant the unbound Erythrina Cristagalli lectin and wash the plate thoroughly with PBS. The decanted lectin solution may be stored at 4°C with 0.05% sodium azide and reused for further plating. 

3. Add 1 x 107 lymphocytes in 4 ml of RPMI-1640 or PBS for 1 h at 4°C. 

4. After collecting the unbound cells (enriched NK fraction), the bound cells can be removed by the addition of 200 mM lactose, 0.5 mM EDTA. 

5. Both the unbound and bound cells can be washed several times with RPMI or PBS containing 200 mM lactose to remove any trace amounts of lectin which might be present.

Especificações

Product Name

Erythrina Cristagalli Lectin (ECL)

Sinónimos

Lectin from Erythrina Cristagalli | ECA

Grau

BioReagent, Native

Especificações e pureza

BioReagent, Native

Conjugação

Unconjugated

Peso molecular previsto

54 kDa

Tipo de molécula

Protein

Product Line

Produtos biológicos

Armazenamento e envio

Reconstituição

Reconstitute in 10 mM HEPES buffered saline, 0.1 mM CaCl₂, pH 8.5. For preserving solutions stored at 4 ºC, 0.08% sodium azide can be used.

Condições de armazenamento de armazenamento

Store at 2-8°C

Enviado em

Wet ice

Estabilidade e armazenamento

Store at 2-8℃ long term. Upon reconstitution, it is recommended to aliquot.