Log In Sign up
The largest database of trusted experimental protocols

Mega 10

Manufactured by Merck Group
Visit Supplier

MEGA-10 is a lab equipment product from Merck Group. It is a non-ionic detergent used for solubilizing and stabilizing proteins and membranes. The core function of MEGA-10 is to serve as a surfactant in various biochemical applications.

Automatically generated - may contain errors

Lab products found in correlation

Quil a saponin, by InvivoGen (4 mentions) Dppc 1 2 dipalmitoyl sn glycero 3 phsophocholine, by Avanti Polar Lipids (2 mentions) Monophosphoryl lipid a, by Avanti Polar Lipids (2 mentions) Mega 10, by Avanti Polar Lipids (2 mentions) Sodium dodecyl sulfate (sds), by Merck Group (1 mentions) Limulus amebocyte lysate assay, by Lonza (1 mentions) Amicon ultra centrifugal filter, by Merck Group (1 mentions) Quil a, by InvivoGen (1 mentions) Cholesterol, by Merck Group (1 mentions)

Close spelling variants of this product

MEGA-10 detergent

7 protocols using mega 10

1

Preparation of Surfactant Solutions

Check if the same lab product or an alternative is used in the 5 most similar protocols
NaBr, MEGA-10, S10S, SDS and DTAB (all from Sigma) were dissolved at a concentration of 173 mM in DI water. Mixtures were sonicated for 1 h at 40 °C for complete dissolution. In most experiments, 6 ml of these solutions were added to 400 ml of DI water using an automatic pipette, bringing the total concentration to 2.6 mM.
Cho H.J., Mizerak J.P, & Wang E.N. (2015). Turning bubbles on and off during boiling using charged surfactants. Nature Communications, 6, 8599.
+ Open protocol
+ Expand
2

Purification and Inactivation of Vaccine Viruses

Check if the same lab product or an alternative is used in the 5 most similar protocols
Vaccine seed viruses were plaque purified on MDCK cells and inoculated into the allantoic fluid of 11-day-old embryonated chicken eggs. Upon incubation at 37°C for 48 h, the allantoic fluid was harvested, cleared by centrifugation at 3,000 rpm (SW 32 Ti; Beckman Counter) for 10 min, and then concentrated by 2 rounds of ultracentrifugation at 27,000 rpm (SW 32 Ti; Beckman Counter) for 2 h at 4°C on 60% (wt/vol) sucrose cushions. The concentrated virus was purified on a 20 to 60% (wt/vol) sucrose gradient at 27,000 rpm (SW 32 Ti; Beckman Counter) overnight at 4°C. The next morning, the virus band was extracted and pelleted by ultracentrifugation at 27,000 rpm (SW 32 Ti; Beckman Counter) at 4°C for 2 h. The resulting pellet was resuspended in 2% Mega10 (Sigma-Aldrich) and incubated for 1 h at 37°C to allow the disruption of the virus by the detergent. The split virus was inactivated with 0.01% formalin solution for 3 days and dialyzed against PBS. The resulting split-inactivated virus preparations were stored in small aliquots at −80°C. Complete viral inactivation was confirmed by three passages on MDCK cells. Levels of endotoxin were determined using the Pierce LAL chromogenic endotoxin quantitation kit according to the instructions from the manufacturer (Thermo Fisher Scientific).
Rosu M.E., Kok A., Bestebroer T.M., de Meulder D., Verveer E.P., Pronk M.R., Dekker L.J., Luider T.M., Richard M., van den Brand J.M., Fouchier R.A, & Herfst S. (2022). Contribution of Neuraminidase to the Efficacy of Seasonal Split Influenza Vaccines in the Ferret Model. Journal of Virology, 96(6), e01959-21.
+ Open protocol
+ Expand
3

Saponin-Based Nanoparticle Adjuvant Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols
The adjuvant used for all the described studies was a ISCOM-like
saponin nanoparticle comprised of self-assembled cholesterol phospholipid,
and quillaja saponin prepared as previously described (Plotkin, 2010 (link)). Briefly, 10 mg each of
cholesterol (Avanti Polar Lipids) and DPPC (Avanti Polar Lipids) were
dissolved separately in 20% MEGA-10 (Sigma-Aldrich) detergent at a final
concentration of 20 mg/mL and 50 mg Quil-A saponin (InvivoGen) was dissolved
in MilliQ H2O at a final concentration of 100 mg/mL. Next, DPPC
solution was added to cholesterol followed by addition of Quil-A saponin in
rapid succession and the volume was brought up with PBS for a final
concentration of 1 mg/mL cholesterol and 2% MEGA-10. The solution was
allowed to equilibrate at 25°C overnight, followed by 5 days of
dialysis against PBS using a 10k MWCO membrane. The adjuvant solution was
filter sterilized using a 0.2 μm Supor syringe filter, concentrated
using 50k MWCO Centricon filters, and further purified by FPLC using a
Sephacryl S-500 HR size exclusion column. Each adjuvant batch was finally
characterized by negative stain transmission electron microscopy (TEM) and
dynamic light scattering (DLS) to confirm uniform morphology and size and
validated for low endotoxin content by Limulus Amebocyte Lysate assay
(Lonza). Final adjuvant concentration was determined by cholesterol
quantification (Sigma-Aldrich).
Cirelli K.M., Carnathan D.G., Nogal B., Martin J.T., Rodriguez O.L., Upadhyay A.A., Enemuo C.A., Gebru E.H., Choe Y., Viviano F., Nakao C., Pauthner M.G., Reiss S., Cottrell C.A., Smith M.L., Bastidas R., Gibson W., Wolabaugh A.N., Melo M.B., Cosette B., Kumar V., Patel N.B., Tokatlian T., Menis S., Kulp D.W., Burton D.R., Murrell B., Schief W.R., Bosinger S.E., Ward A.B., Watson C.T., Silvestri G., Irvine D.J, & Crotty S. (2019). Slow delivery immunization enhances HIV neutralizing antibody and germinal center responses via modulation of immunodominance. Cell, 177(5), 1153-1171.e28.
+ Open protocol
+ Expand
4

Preparation and Purification of SMNP Adjuvant

Check if the same lab product or an alternative is used in the 5 most similar protocols
SMNP adjuvant was prepared as previously described (27 ). Briefly, solutions at 20 mg/ml were prepared of cholesterol (Avanti Polar Lipids, catalog no. 700000), dipalmitoylphosphatidylcholine (DPPC; Avanti Polar Lipids, catalog no. 850355), and PHAD MPLA (Avanti Polar Lipids, catalog no. 699800P) in 20% N-decanoyl-N-methylglucamine (MEGA-10) (Sigma-Aldrich, catalog no. D6277) detergent. Quil-A saponin (InvivoGen, catalog no. vac-quil) was dissolved in Milli-Q water at a final concentration of 100 mg/ml. These were mixed at a mass ratio of 10:2:1:1 (Quil-A:chol:DPPC:MPLA) and diluted in PBS to a final cholesterol concentration of 1 mg/ml. The solution was equilibrated overnight at 25°C and then dialyzed against PBS using a 10-kDa MWCO cassette. The adjuvant was then sterile filtered, concentrated using Amicon Ultra centrifugal filters (50-kDa MWCO; MilliporeSigma, catalog no. UFC505096), and purified by FPLC using a Sephacryl S-500 HR size exclusion column. SMNP labeled with Cy7 was prepared as described incorporating 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(Cyanine 7) (Avanti Polar Lipids, catalog no. 810347) in place of 10 mole percent of the MPLA.
Rodrigues K.A., Rodriguez-Aponte S.A., Dalvie N.C., Lee J.H., Abraham W., Carnathan D.G., Jimenez L.E., Ngo J.T., Chang J.Y., Zhang Z., Yu J., Chang A., Nakao C., Goodwin B., Naranjo C.A., Zhang L., Silva M., Barouch D.H., Silvestri G., Crotty S., Love J.C, & Irvine D.J. (2021). Phosphate-mediated coanchoring of RBD immunogens and molecular adjuvants to alum potentiates humoral immunity against SARS-CoV-2. Science Advances, 7(50), eabj6538.
+ Open protocol
+ Expand
5

Preparation of ISCOM with rEtpE-C

Check if the same lab product or an alternative is used in the 5 most similar protocols
ISCOM with rEtpE-C was prepared as described previously (50 ) with slight modifications. Briefly, purified EtpE-C (1 ml; 0.5 to 1 mg/ml) that had been eluted from the affinity-purification column with 8 M urea was dissolved in 1 ml of 20% Mega 10 (N-decanoyl-N-methylglucamine; Sigma) and mixed with 100 μl of 1 mg/ml of cholesterol (Sigma), 1 mg/ml phospholipid (l-α-phosphatidylcholine), and 50 μl of 100 mg/ml Quil A (InvivoGen, San Diego, CA). The mix was rotated for 2 h at room temperature and sonicated in a water bath three times for 15 min each. The mix was placed in dialysis tubing (10 kDa MW cutoff; Spectrum, New Brunswick, NJ) and dialyzed against PBS for 24 h at room temperature and dialyzed again for 24 h at 4°C. ISCOM with PBS was prepared as a negative control, and ISCOM preparations were stored at –80°C or used directly for vaccinations.
Budachetri K., Teymournejad O., Lin M., Yan Q., Mestres-Villanueva M., Brock G.N, & Rikihisa Y. (2020). An Entry-Triggering Protein of Ehrlichia Is a New Vaccine Candidate against Tick-Borne Human Monocytic Ehrlichiosis. mBio, 11(4), e00895-20.
+ Open protocol
+ Expand
6

Preparation and Purification of Saponin-based Adjuvant

Check if the same lab product or an alternative is used in the 5 most similar protocols
SMNP adjuvant was prepared as described (77 ). Cholesterol (20 mg/ml; Avanti Polar Lipids, catalog no. 700000) and DPPC (1,2-dipalmitoyl-sn-glycero-3-phsophocholine: 20 mg/ml; Avanti Polar Lipids, catalog no. 850355) were dissolved in Milli-Q water containing 20% w/v MEGA-10 (Sigma-Aldrich, D6277) detergent at 60°C. Monophosphoryl lipid A (10 mg/ml; Avanti Polar Lipids, catalog no. 699800P) was dissolved in 20% w/v MEGA-10 at 37°C. Quil-A saponin (InvivoGen; vac-quil) was dissolved in Milli-Q water at a final concentration of 100 mg/ml at 37°C. All components were then mixed at a mass ratio of 10:10:2.5:1 [Quil-A:cholesterol:DPPC:MPLA (Monophosphoryl lipid A)] at 60°C followed by dilution with phosphate-buffered saline (PBS) to a final concentration of cholesterol (1 mg/ml). The solution was allowed to equilibrate over night at room temperature, followed by dialysis against PBS using a 10k molecular weight cutoff (MWCO) membrane. The adjuvant solution was then sterile filtered, concentrated using 50k MWCO centricon spin filters, and further purified by fast protein liquid chromatography using a Sephacryl S-500 HR size exclusion column. Cited concentrations of SMNP refer to the concentration of saponin injected.
He W.T., Yuan M., Callaghan S., Musharrafieh R., Song G., Silva M., Beutler N., Lee W.H., Yong P., Torres J.L., Melo M., Zhou P., Zhao F., Zhu X., Peng L., Huang D., Anzanello F., Ricketts J., Parren M., Garcia E., Ferguson M., Rinaldi W., Rawlings S.A., Nemazee D., Smith D.M., Briney B., Safonova Y., Rogers T.F., Dan J.M., Zhang Z., Weiskopf D., Sette A., Crotty S., Irvine D.J., Ward A.B., Wilson I.A., Burton D.R, & Andrabi R. (2022). Broadly neutralizing antibodies to SARS-related viruses can be readily induced in rhesus macaques. Science translational medicine, 14(657), eabl9605.
+ Open protocol
+ Expand
7

Preparation and Purification of Saponin-based Adjuvant

Check if the same lab product or an alternative is used in the 5 most similar protocols
SMNP adjuvant was prepared as described (77 ). Cholesterol (20 mg/ml; Avanti Polar Lipids, catalog no. 700000) and DPPC (1,2-dipalmitoyl-sn-glycero-3-phsophocholine: 20 mg/ml; Avanti Polar Lipids, catalog no. 850355) were dissolved in Milli-Q water containing 20% w/v MEGA-10 (Sigma-Aldrich, D6277) detergent at 60°C. Monophosphoryl lipid A (10 mg/ml; Avanti Polar Lipids, catalog no. 699800P) was dissolved in 20% w/v MEGA-10 at 37°C. Quil-A saponin (InvivoGen; vac-quil) was dissolved in Milli-Q water at a final concentration of 100 mg/ml at 37°C. All components were then mixed at a mass ratio of 10:10:2.5:1 [Quil-A:cholesterol:DPPC:MPLA (Monophosphoryl lipid A)] at 60°C followed by dilution with phosphate-buffered saline (PBS) to a final concentration of cholesterol (1 mg/ml). The solution was allowed to equilibrate over night at room temperature, followed by dialysis against PBS using a 10k molecular weight cutoff (MWCO) membrane. The adjuvant solution was then sterile filtered, concentrated using 50k MWCO centricon spin filters, and further purified by fast protein liquid chromatography using a Sephacryl S-500 HR size exclusion column. Cited concentrations of SMNP refer to the concentration of saponin injected.
He W.T., Yuan M., Callaghan S., Musharrafieh R., Song G., Silva M., Beutler N., Lee W.H., Yong P., Torres J.L., Melo M., Zhou P., Zhao F., Zhu X., Peng L., Huang D., Anzanello F., Ricketts J., Parren M., Garcia E., Ferguson M., Rinaldi W., Rawlings S.A., Nemazee D., Smith D.M., Briney B., Safonova Y., Rogers T.F., Dan J.M., Zhang Z., Weiskopf D., Sette A., Crotty S., Irvine D.J., Ward A.B., Wilson I.A., Burton D.R, & Andrabi R. (2022). Broadly neutralizing antibodies to SARS-related viruses can be readily induced in rhesus macaques. Science translational medicine, 14(657), eabl9605.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!