HeLa cells were lysed with RIPA lysis buffer (150 mM NaCl, 50 mM Tris, pH 7.4, 1 mM EDTA, and 0.5% Nonidet P-40), and cell lysates (200 μg of protein) were incubated with agarose-conjugated sWGA beads (Vector Laboratories, Burlingame, CA, USA) overnight at 4°C. For control samples, the inhibitory monosaccharide GlcNAc was added during the sWGA-lectin-affinity purification. Precipitates were washed three times with RIPA buffer and proteins were eluted by boiling in SDS sample buffer. Lysates were then analysed by western blot.
Agarose conjugated swga beads
Manufactured by Vector Laboratories
Sourced in United States
Agarose-conjugated sWGA beads are a laboratory product used for the purification and isolation of specific biomolecules. The beads are composed of agarose, a polysaccharide derived from seaweed, and are conjugated with sWGA, a wheat germ agglutinin lectin that binds to N-acetylglucosamine residues. These beads can be used in various biochemical and molecular biology applications that require the separation and enrichment of targeted molecules.
Automatically generated - may contain errors
6 protocols using agarose conjugated swga beads
1
Glycoprotein Enrichment from HeLa Cells
2
Glycoprotein Enrichment from Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa and HaCaT cells were lysed with RIPA lysis buffer (150 mM NaCl, 50 mM Tris, pH 7.4, 1 mM EDTA, 0.5% Nonidet P-40), and the cell lysates (∼200 µg protein) were incubated with agarose-conjugated sWGA beads (Vector Laboratories, Burlingame, CA, USA) overnight at 4°C. For the control, the inhibitory monosaccharide GlcNAc was added during the sWGA-lectin-affinity purification. Precipitates were washed three times with lysis buffer, and the proteins were eluted by boiling in SDS sample buffer.
3
Selective Lectin-Affinity Purification of GlcNAc Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa cells were lysed with RIPA lysis buffer (150mM NaCl, 50mM Tris, pH 7.4, 1mM EDTA, 0.5% Nonidet P-40) and cell lysates(~100 μg of protein) was incubated with agarose-conjugated sWGA beads (Vector Laboratories, Burlingame, CA) overnight at 4°C. For control, the inhibitory monosaccharide GlcNAc was added during sWGA-lectin-affinity purification. Precipitates were washed three times with lysis buffer and proteins were eluted by boiling in SDS sample buffer.
4
Wheat Germ Agglutinin Affinity Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in RIPA buffer (Cat # P0013B, Beyotime Biotechnology, Shanghai, China) containing 50 mM Tris–HCl (pH 7.4), 150 mM NaCl, 1% NP-40, 0.1% SDS. Succinylated wheat germ agglutinin (sWGA)-conjugated agarose beads (Cat # AL-1023S-2, Vector Laboratories) were cleaned in washing buffer containing 1/3 RIPA buffer (Cat # P0013B, Beyotime) and 2/3 PBS plus PhosSTOP (Cat # 04906845001, Roche) and EDTA-free protease inhibitors (Cat # 04693159001, Roche). Pre-cleaned lysates were incubated overnight with sWGA-conjugated agarose beads in 4℃. Precipitated complexes were eluted with washing buffer. Loading buffer (5X) (Cat # P1040, Solarbio) and precipitated complexes were mixed and denatured at 100 °C for 5 min. Precipitated complexes were immunoblotted with the indicated antibodies.
5
Isolation of N-Linked Glycoproteins from Hepatic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hepatic cells and liver tissues were lysed in a buffer containing 125 mM NaCl; 50 mM Tris, pH 7.4; 5 mM EDTA; and 0.1% NP-40. The lysate was denatured in glycoprotein-denaturing buffer and digested with PNGase F (P0704S, New England Biolabs, Ipswitch, MA, USA) to remove N-linked glycoproteins. Pre-cleared lysates were incubated overnight with sWGA-conjugated agarose beads (Vector Laboratories, Burlingame, CA, USA). Precipitated complexes were eluted and immunoblotted with the indicated antibodies.
6
Isolation and Analysis of N-Glycoproteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Liver tissues or hepatic cells were lysed in Lysis 125 buffer (50 mM Tris, pH 7.4, 125 mM NaCl, 5 mM EDTA, 5 mM EGTA, 0.1% Nonidet P-40, 50 mM NaF, 1 mM PMSF, and 1× Proteinase Inhibitor Cocktail (Roche)). The supernatant was denatured in glycoprotein denaturing buffer and digested with PNGase (P0704S; New England Biolabs, USA) to remove N-linked glycoproteins. Pre-cleared supernatant was incubated with succinylated wheat-germ agglutinin (sWGA)-conjugated agarose beads (Vector Laboratories, Burlingame, CA) overnight at 4 °C. Precipitated complexes were eluted and immunoblotted with anti-KAT5 antibodies.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!