0.45nm nitrocellulose membrane

Manufactured by Bio-Rad

The 0.45nm nitrocellulose membrane is a laboratory filtration product used to separate and retain small biomolecules, such as proteins and nucleic acids, during various analytical and preparative processes. It is a thin, porous membrane with a nominal pore size of 0.45 micrometers, which allows for the efficient capture and retention of the target molecules while permitting the passage of smaller components.

Automatically generated - may contain errors

Lab products found in correlation

Chemidoc xrs system, by Bio-Rad (2 mentions) Anti akt antibody 9272, by Cell Signaling Technology (1 mentions) Image labtm software, by Bio-Rad (1 mentions) Ecl anti rabbit igg hrp, by GE Healthcare (1 mentions) Precision plus protein standard, by Bio-Rad (1 mentions) Ripa buffer, by Thermo Fisher Scientific (1 mentions) Anti b actin mouse monoclonal antibody, by Merck Group (1 mentions) Image lab software, by Bio-Rad (1 mentions)

3 protocols using 0.45nm nitrocellulose membrane

Western Blot Analysis of pAKT and AKT

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total protein lysates (20ng) of BAL samples or THP-1 transfected cells were lysed with RIPA lysis buffer supplemented with protease and phosphatase inhibitors for phosphoprotein and total protein analyses. Lysates were separated in 12% SDS-PAGE, transferred to 0.45nm nitrocellulose membrane (Biorad), followed by Western blot detection of pAKT^ser473, total AKT and b-actin, with anti-pAKT^ser473 antibody #9271, (Cell Signaling), anti-AKT antibody # 9272 (Cell Signaling) and anti-b-actin monoclonal antibody (Chemicon). Appropriate HRP conjugated secondary antibodies (Chemicon) were used and immunodetection was performed with enhanced chemiluminescence reagent Luminata^TM (Millipore). Bands were visualised with the ChemiDoc XRS+ system (Biorad) and densitometry analyses were performed using Image Lab ^TM software (Biorad).

Tsitoura E., Wells A.U., Karagiannis K., Lasithiotaki I., Vasarmidi E., Bibaki E., Koutoulaki C., Sato H., Spandidos D.A., Siafakas N.M., Sourvinos G, & Antoniou K.M. (2016). MiR-185/AKT and miR-29a/Collagen 1a pathways are activated in IPF BAL cells. Oncotarget, 7(46), 74569-74581.

+ Open protocol

+ Expand

Western Blot Analysis of O-GlcNAc Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

All chemicals and antibodies were from Sigma Aldrich (St. Louis, MO) unless noted otherwise. Precision Plus Protein Standard, 4–15% or 8–16% Criterion TGX Precast gels, 10x TGS running buffer, and 0.45 nm nitrocellulose membrane were from Bio-Rad (Hercules, CA) and 10x ReBlot Plus Strong Stripping Antibody Solution was from Millipore (Temecula, CA). Primary antibodies used in this study were monoclonal anti-O-GlcNAc antibody (clone CTD110.6), rabbit anti-actin antibody, monoclonal anti-β-actin antibody, monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody. Anti-OGT antibody clone AL28 was kindly provided by S. Arnold, Johns Hopkins University, Baltimore, MD, and anti-OGA antibody was kindly provided by G. Crawford, Mercer University, Macon, GA. Secondary antibodies used were anti-mouse IgG/IgM horseradish peroxidase (HRP) (Millipore, Temecula, CA), ECL^™ anti-rabbit IgG HRP, ECL^™ Plex goat-anti-mouse IgG, Cy^™5, ECL^™ Plex goat-anti-rabbit IgG, Cy^™5 (all GE Healthcare, Pittsburgh, PA) and anti-rabbit IgG alkaline phosphatase.

Förster S., Welleford A.S., Triplett J.C., Sultana R., Schmitz B, & Butterfield D.A. (2014). Increased O-GlcNAc Levels Correlate with Decreased O-GlcNAcase Levels in Alzheimer Disease Brain. Biochimica et biophysica acta, 1842(9), 1333-1339.

+ Open protocol

+ Expand

Mitochondrial Protein Profiling in BAL

Check if the same lab product or an alternative is used in the 5 most similar protocols

1–1.5 million cells were centrifuged and cell pellets were homogenised in RIPA buffer (Invitrogen) containing protease and phosphatase inhibitors, Pierce), followed by storage at -80^οC. 40–60 μg Total protein lysates of BAL samples were separated in 12% SDS-PAGE, transferred to 0.45 nm nitrocellulose membrane (Biorad), followed by detection of PINK1[anti-PINK1, mouse monoclonal antibody, (Novus Biologicals)], TOMM20 [anti-TOMM20, rabbit polyclonal (Abnova)], p62 [anti-p62 mouse monoclonal antibody (MBL)], LC3 [anti-LC3 mouse monoclonal antibody (Abgent)] and b-actin [anti-b-actin mouse monoclonal antibody (Sigma)]. Appropriate anti-mouse HRP conjugated secondary antibody (Chemicon) was used and immunodetection was performed with enhanced chemiluminescence reagent Luminata™ (Millipore). Bands were visualised with the ChemiDocXRS+ system (Biorad) and densitometry analyses were performed using Image Lab™ software (Biorad).

Tsitoura E., Vasarmidi E., Bibaki E., Trachalaki A., Koutoulaki C., Papastratigakis G., Papadogiorgaki S., Chalepakis G., Tzanakis N, & Antoniou K.M. (2019). Accumulation of damaged mitochondria in alveolar macrophages with reduced OXPHOS related gene expression in IPF. Respiratory Research, 20, 264.

+ 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?

Revolutionizing how scientists
search and build protocols!