Log In Sign up
The largest database of trusted experimental protocols

Immobilon classico

Manufactured by Merck Group
Visit Supplier

Immobilon Classico is a lab equipment product manufactured by Merck Group. It is designed for the immobilization of biomolecules, such as proteins, enzymes, or antibodies, onto solid supports. The core function of Immobilon Classico is to facilitate the attachment and stabilization of these biomolecules for various analytical and research applications.

Automatically generated - may contain errors

Lab products found in correlation

Crescendo western hrp substrate, by Merck Group (4 mentions) Stripping solution, by Bio-Rad (2 mentions) Phosphatase inhibitor, by Merck Group (2 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (2 mentions) Micro bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Protease inhibitor, by Merck Group (2 mentions) Trans blot turbo transfer system, by Bio-Rad (2 mentions) Anti actin, by Merck Group (2 mentions) Bis tris acrylamide, by Bio-Rad (2 mentions) Horseradish peroxidase hrp conjugated mouse monoclonal anti flag, by Merck Group (2 mentions) Hrp conjugated mouse monoclonal anti gapdh, by Proteintech (2 mentions) Anti crkrs cdk12, by Novus Biologicals (2 mentions) Anti cdc2l5 cdk13, by Novus Biologicals (2 mentions) Anti cyck, by Abcam (2 mentions) Anti ha antibody, by Merck Group (2 mentions) Laemmli sample buffer, by Bio-Rad (2 mentions) Protease inhibitor cocktail, by Merck Group (2 mentions) Nitrocellulose membrane, by Bio-Rad (2 mentions) Precision plus protein kaleidoscope molecular, by Bio-Rad (1 mentions) A3854, by Merck Group (1 mentions)

5 protocols using immobilon classico

1

Whole Cell Lysate Preparation and Immunoblotting

Check if the same lab product or an alternative is used in the 5 most similar protocols
Radioimmunoprecipitation (RIPA) buffer supplemented with phenylmethane-sulfonylfluoride (PMSF) (Sigma), protease inhibitor (Sigma), and phosphatase inhibitor (Sigma) was used to make whole cell lysates as previously described28 . Protein concentrations were determined using a Micro BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA), and samples were loaded into sodium dodecyl sulfate polyacrylamide (SDS-PAGE) gels adjacent to the Precision Plus Protein Kaleidoscope molecular weight marker (Bio-Rad, Hercules, CA). Proteins were separated via electrophoresis and gels transferred to blotting membrane using Trans-Blot Turbo Transfer System (Bio-Rad) per manufacturer’s protocol. Antibodies were utilized per manufacturers’ instructions. Immunoblots were developed with Immobilon Classico or Crescendo Western HRP Substrate (EMD Millipore, Millipore Sigma, Burlington, MA). Blots were stripped with stripping solution (Bio-Rad) at 65 °C for 20 minutes and then re-probed with selected antibodies. Equal protein loading was confirmed using β-actin.
Stafman L.L., Williams A.P., Marayati R., Aye J.M., Markert H.R., Garner E.F., Quinn C.H., Lallani S.B., Stewart J.E., Yoon K.J., Whelan K, & Beierle E.A. (2019). Focal Adhesion Kinase Inhibition Contributes to Tumor Cell Survival and Motility in Neuroblastoma Patient-Derived Xenografts. Scientific Reports, 9, 13259.
+ Open protocol
+ Expand
2

Whole-Cell Lysate Isolation and Protein Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole-cell lysates were isolated using radioimmunoprecipitation (RIPA) buffer supplemented with protease inhibitor (Sigma), phosphatase inhibitor (Sigma), and phenylmethane-sulfonylfluoride (PMSF) (Sigma) as previously described (15 (link)). Protein concentrations were determined using a Micro BCA Protein Assay Kit (Thermo Fisher Scientific Inc.), and samples were separated by electrophoresis on sodium dodecyl sulfate polyacrylamide (SDS-PAGE) gels. The Precision Plus Protein Kaleidoscope molecular weight marker (Bio-Rad, Hercules, CA) was utilized to confirm expected size of target proteins. Gel transfer to blotting membrane was completed using Trans-Blot Turbo Transfer System (Bio-Rad) per manufacturer’s protocol. Antibodies were utilized according to the manufacturers’ recommendations. Immunoblots were developed with Immobilon Classico or Crescendo Western HRP Substrate (EMD Millipore, Millipore Sigma, Burlington, MA). Blots were stripped with stripping solution (Bio-Rad) at 65°C for 20 minutes and then re-probed with selected antibodies. Equal protein loading between samples was confirmed using β-actin as an internal control.
Marayati R., Williams A.P., Bownes L.V., Quinn C.H., Stewart J.E., Mroczek-Musulman E., Atigadda V.R, & Beierle E.A. (2020). Novel Retinoic Acid Derivative Induces Differentiation and Growth Arrest in Neuroblastoma. Journal of pediatric surgery, 55(6), 1072-1080.
+ Open protocol
+ Expand
3

SDS-PAGE and Immunoblotting of NNMT

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were prepared as previously described [Drabarek et al., 2012; (link)Dymkowska et al., 2014] (link). Proteins were separated by polyacrylamide gel electrophoresis (PAGE) under denaturing conditions in the presence of 0.1% sodium dodecyl sulphate (SDS, BioShop) [Laemmli, 1970] (link). After transferring to the PVDF membrane (Millipore) NNMT protein was detected with the use of a specific primary antibody (Santa Cruz Biotechnology, sc-376048); anti-rabbit and anti-mouse secondary antibodies conjugated with horseradish peroxidase (HRP) were obtained from Abcam. For HRP detection (Fusion FX, Vilber Lourmat) chemiluminescent substrate Immobilon Classico (Merck Millipore) was used. The optical density of bands corresponding to defined proteins was determined densitometrically with the BIO-1D software (Vilber Lourmat) and expressed in relation to β-actin used as a loading control (Sigma, A-3854).
Stępińska O., Dymkowska D., Mateuszuk Ł., & Zabłocki K. (2021). Effect of lipopolysaccharide (LPS) on HAEC cells. Does nicotinamide N-methyltransferase sensitize HAEC cells to LPS?.
+ Open protocol
+ Expand
4

Western Blot Analysis of Cellular Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed with 1% NP-40 in PBS containing protease inhibitor cocktail (Sigma). After removal of nuclei by brief centrifugation, cytosolic fraction was collected and mixed with 4x Laemmli Sample Buffer (Bio-Rad). Proteins were separated in 10% Bis-Tris acrylamide gels, blotted on to nitrocellulose membrane (Bio-Rad), and detected by antibodies. Mouse monoclonal anti-Nef was described previously (Zheng et al., 2003 (link)). Horseradish peroxidase (HRP)-conjugated mouse monoclonal anti-FLAG, anti-HA antibodies, and anti-actin were purchased from Sigma; HRP-conjugated mouse monoclonal anti-GAPDH was purchased from Proteintech; rabbit polyclonal anti-CrkRS (CDK12) and anti-CDC2L5 (CDK13) were purchased from Novus; rabbit polyclonal anti-CycK was purchased from Abcam; HRP-conjugated anti-human, -rabbit, and -mouse immunoglobulin G secondary antibodies were purchased from Thermo Fisher. Immobilon Classico and Crescendo Western HRP Substrate was purchased from MilliporeSigma.
Chai Q., Li S., Collins M.K., Li R., Ahmad I., Johnson S.F., Frabutt D.A., Yang Z., Shen X., Sun L., Hu J., Hultquist J.F., Peterlin B.M, & Zheng Y.H. (2021). HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity. Cell reports, 36(6), 109514.
+ Open protocol
+ Expand
5

Western Blot Analysis of Cellular Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed with 1% NP-40 in PBS containing protease inhibitor cocktail (Sigma). After removal of nuclei by brief centrifugation, cytosolic fraction was collected and mixed with 4x Laemmli Sample Buffer (Bio-Rad). Proteins were separated in 10% Bis-Tris acrylamide gels, blotted on to nitrocellulose membrane (Bio-Rad), and detected by antibodies. Mouse monoclonal anti-Nef was described previously (Zheng et al., 2003 (link)). Horseradish peroxidase (HRP)-conjugated mouse monoclonal anti-FLAG, anti-HA antibodies, and anti-actin were purchased from Sigma; HRP-conjugated mouse monoclonal anti-GAPDH was purchased from Proteintech; rabbit polyclonal anti-CrkRS (CDK12) and anti-CDC2L5 (CDK13) were purchased from Novus; rabbit polyclonal anti-CycK was purchased from Abcam; HRP-conjugated anti-human, -rabbit, and -mouse immunoglobulin G secondary antibodies were purchased from Thermo Fisher. Immobilon Classico and Crescendo Western HRP Substrate was purchased from MilliporeSigma.
Chai Q., Li S., Collins M.K., Li R., Ahmad I., Johnson S.F., Frabutt D.A., Yang Z., Shen X., Sun L., Hu J., Hultquist J.F., Peterlin B.M, & Zheng Y.H. (2021). HIV-1 Nef interacts with the cyclin K/CDK13 complex to antagonize SERINC5 for optimal viral infectivity. Cell reports, 36(6), 109514.
+ 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!