Log In Sign up
The largest database of trusted experimental protocols

Nupage 4 12 bis tris gradient minigels

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in Canada, United States

The NuPAGE 4–12% Bis-Tris gradient minigels are precast polyacrylamide gels designed for protein separation and analysis. The gels feature a gradient of acrylamide concentrations, allowing for the separation of a wide range of protein molecular weights in a single run.

Automatically generated - may contain errors

Lab products found in correlation

Odyssey infrared imaging system, by LI COR (4 mentions) Immobilon polyvinylidene difluoride membrane, by Thermo Fisher Scientific (2 mentions) Pvdf membrane, by Thermo Fisher Scientific (1 mentions) Anti gfp monoclonal antibody, by Takara Bio (1 mentions) Anti mouse irdye 680, by LI COR (1 mentions) Anti mouse irdye 800, by LI COR (1 mentions) Ripa buffer, by Merck Group (1 mentions) Polyvinylidene fluoride (pvdf), by Thermo Fisher Scientific (1 mentions) Anti flag m2 monoclonal antibody, by Merck Group (1 mentions) Cell surface protein isolation kit, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

4-12 Bis-Tris minigels NuPAGE 4–12% Bis-Tris NuPAGE 12% Bis-Tris 12% Bis-Tris minigels NuPAGE Bis-Tris NuPAGE minigels 4–12% gradient minigels NuPAGE

4 protocols using nupage 4 12 bis tris gradient minigels

1

Mitochondrial Protein Isolation and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mitochondria were isolated from HepG2 stable cells and the total protein was isolated by using RIPA buffer (Sigma) followed by sonication. The total protein and mitochondrial protein (~60 μg) were resolved in NuPAGE 4–12% Bis-Tris gradient minigels (Invitrogen) followed by transfer onto PVDF membrane (Fisher Scientific), and subsequently membranes were incubated with anti-GFP monoclonal antibody (Clontech, CA) and anti-pyruvate dehydrogenase (PDH) monoclonal antibodies (Abcam, MA). The detection of specific protein bands were performed by incubating with anti-mouse IRDye-800 and anti-mouse IRDye-680 secondary antibodies (LI-COR Bioscience, Lincoln, NE) at 1:30,000 dilutions sequentially. Finally, the bands were visualized using the Odyssey infrared imaging system (LI-COR Bioscience) [30 (link)]. The density of individual band was determined by using odyssey application software (version 3.0).
Sabui S., Subramanian V.S., Kapadia R, & Said H.M. (2016). Structure - function characterization of the human mitochondrial thiamin pyrophosphate transporter (hMTPPT; SLC25A19): important roles for Ile33, Ser34, Asp37, His137 and Lys291. Biochimica et biophysica acta, 1858(8), 1883-1890.
+ Open protocol
+ Expand
2

Quantitative Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were prepared from vitamin-C-treated and untreated BEAS-2B cells. Sixty micrograms of protein samples were separated in NuPAGE 4–12% Bis-Tris gradient minigels (Invitrogen, Carlsbad, CA, USA), then transferred onto Immobilon polyvinylidene difluoride membrane (Fisher Scientific, Chino, CA, USA). The well-characterized anti-SVCT-2 (1:200 dilutions) and β-actin (1:2000 dilutions) antibodies were used as primary antibodies. The anti-rabbit IRDye-800 and anti-mouse IRDye-680 (both at 1:30,000 dilutions) were used as secondary antibodies. The immunoreactive bands were quantified using the Odyssey infrared imaging system (LI-COR Biosciences, Lincoln, NE, USA).
Teafatiller T., Agrawal S., De Robles G., Rahmatpanah F., Subramanian V.S, & Agrawal A. (2021). Vitamin C Enhances Antiviral Functions of Lung Epithelial Cells. Biomolecules, 11(8), 1148.
+ Open protocol
+ Expand
3

Immunoblotting Analysis of hRFVT Transporters

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were isolated from RF deficient and OS NCM460 cells and separated in NuPAGE 4–12% Bis-Tris gradient minigels (Invitrogen), transferred onto immobilon polyvinylidene difluoride membrane (PVDF) (Fisher Scientific). The anti-hRFVT-1, -2, -3, Sp1 and anti-β-actin antibodies were used as primary antibodies. The secondary antibodies used were anti-rabbit IRDye-800 and anti-mouse IRDye-680 (both at 1:30,000 dilutions). The specific immunoreactive bands were detected using the Odyssey infrared imaging system (LI-COR Bioscience, Lincoln, NE) and their densities were quantified using the LI-COR software.
Subramanian V.S., Ghosal A., Kapadia R., Nabokina S.M, & Said H.M. (2015). Molecular Mechanisms Mediating the Adaptive Regulation of Intestinal Riboflavin Uptake Process. PLoS ONE, 10(6), e0131698.
+ Open protocol
+ Expand
4

Western Blot Analysis of hTPPT Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole cell lysates or plasma membrane proteins isolated with a cell surface protein isolation kit (Pierce Biotechnology, Rockford, IL) were prepared as described [27 ]. Proteins were separated in NuPAGE 4–12% Bis-Tris gradient minigels (Invitrogen), transferred onto immobilon polyvinylidene difluoride membrane (Fisher Scientific), and subjected to Western blot analysis. The primary antibodies were goat polyclonal anti-hTPPT antibodies [CTL4 (D-14) antibody, catalog # sc-68049; Santa Cruz Biotechnology, Santa Cruz, CA] at 1:200 dilution or monoclonal anti-FLAG M2 antibody (Sigma) at 1:1000 dilution. The secondary antibodies were anti-goat IRDye-800 antibody or anti-mouse IRDye-800 antibodies (LI-COR Bioscience, Lincoln, NE) at 1:30,000 dilutions. Immunoreactive bands were visualized using the Odyssey infrared imaging system (LI-COR Bioscience, Lincoln, NE).
Nabokina S.M., Subramanian V.S, & Said H.M. (2016). The human colonic thiamine pyrophosphate transporter (hTPPT) is a glycoprotein and N-linked glycosylation is important for its function. Biochimica et biophysica acta, 1858(4), 866-871.
+ 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!