Nemopilema Nomurai Lectin Production Service

The molecular weight of Nemopilema nomurai lectin (NnL) is 28 kDa. It specifically binds N-acetylneuraminic acid, N-acetyl-D-galactosamine, and N-glycolylneuraminic acid. CD BioGlyco provides NnL production services while helping clients to quickly and accurately analyze the tissue positioning and physiological activity of NnL.

NnL Extraction Service at CD BioGlyco

Our staff selects healthy live Nemopilema nomurai with medium size, smooth surface, and no foreign matter. The tentacles are refrigerated in liquid nitrogen as soon as collection. All manipulations of the tentacles for protein purification are performed on ice and stored at -80°C.

NnL Purification Service at CD BioGlyco

The tissues are homogenized in acetate buffer, then collection the supernatant. We use an ion exchange chromatography column for its first purification. The resulting fractions are dialyzed at 4°C for some time. Dialysis samples are purified a second time by an affinity chromatography column, while the impurity and target fractions are adsorbed using various buffers. Notably, the secondary elution remains slow. Small batches are collected with multiple tubes to increase the purity.

NnL Analysis Service at CD BioGlyco

Further detailed analysis of the NnL makes it very necessary, which is the best way to expand its application and deeply study its expression pattern. We offer the following services:

Hemagglutination and inhibition assay

We use a two-fold serial dilution method to determine hemagglutination activity.
At the same time, the sugar and glycoprotein solutions are serially diluted and added to the lectin solution. After incubation, erythrocyte suspensions are added and inhibition is observed.

Agglutination of bacteria

Collection of cultured bacteria washed several times with buffer solution. Immobilization of bacteria at 4°C for a long time, secondary washing. Mix immobilized bacteria and lectin solution and incubate at room temperature. Observation of aggregation under a light microscope.

SDS-PAGE and amino acid sequencing

Preliminary molecular weight determination is performed using SDS-PAGE to observe the bands.
The lectins are first separated by SDS-PAGE and transferred to biofilm cultures using a western blot. After staining, the assay is performed using a protein sequencer.

cDNA cloning and sequence analysis

We provide lectin cDNA amplification service and amino acid sequence detection service. Including but not limited to signal peptide cleavage sites, isoelectric point detection, structural domain analysis, and lectin protein homology assay.

Fig.1 Schematic diagram of NnL production services. (CD BioGlyco)

Publication

Paper Title: Purification, characterization, and cDNA cloning of a novel lectin from the jellyfish Nemopilema nomurai

Journal: Comparative Biochemistry and Physiology, Part B

IF: 2.495

Published: 2010

Results: For post-purification lectins, a 28 kDa band was detected on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. NnL had the ability to agglutinate horse erythrocytes, gram-positive bacteria, Bacillus subtilis, and gram-negative bacilli Escherichia coli K12. The primary structure of NnL contained part of the fibrinogen-like domain.

Fig.2 SDS-PAGE and agglutination of bacteria by NnL. (Imamichi & Yokoyama, 2010) Fig.2 SDS-PAGE and agglutination of bacteria by NnL. (Imamichi & Yokoyama, 2010)

Applications of NnL

NnL is involved in the innate immune response of Nemopilema nomurai.
Based on binding specificity, NnL is used as a reagent for detecting sugar chains in biochemical and histochemical studies.
NnL has several functions in the immune system and the acquisition of symbionts, which enable it to agglutinate bacteria.

Advantages of Us

Our dual-column elution service ensures high-purity purification.
Our team of professionals provides analytical services customized to the needs of our clients.

CD BioGlyco always puts the client's needs first. Combining multiple biological platforms and trained technicians, we provide professional Lectin Production Services to clients. Please feel free to contact us if you are interested in the Animal Lectin program.

Reference

Imamichi, Y.; Yokoyama, Y. Purification, characterization and cDNA cloning of a novel lectin from the jellyfish Nemopilema nomurai. Comp Biochem Physiol B Biochem Mol Biol. 2010, 156(1): 12-8.

This service is for Research Use Only, not intended for any clinical use.