Log In Sign up
The largest database of trusted experimental protocols

Precast 4 20 polyacrylamide gels

Manufactured by Bio-Rad
Visit Supplier

Precast 4-20% polyacrylamide gels are a type of laboratory equipment used for protein separation and analysis. These gels provide a consistent and uniform matrix for electrophoretic separation of proteins based on their molecular weight. The 4-20% gradient allows for the separation of a wide range of protein sizes.

Automatically generated - may contain errors

Lab products found in correlation

Chemidoc xrs imager, by Bio-Rad (2 mentions) Western ecl substrate, by Bio-Rad (2 mentions) Anti hsp27, by Cell Signaling Technology (2 mentions) Oneblock western fl blocking buffer, by Genesee Scientific (2 mentions) Anti o glcnac rl2, by Thermo Fisher Scientific (2 mentions) Anti αbc, by Cell Signaling Technology (2 mentions) Sequencing grade trypsin, by Roche (1 mentions) Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions) Nitrocellulose blotting membrane, by GE Healthcare (1 mentions)

Spelling variants of this product

Polyacrylamide gel (457 citations) Precast gels (207 citations) 4–20% precast polyacrylamide gel (178 citations) Mini-PROTEAN® TGXTM 4–20% polyacrylamide precast SDS-PAGE gels (2 citations) Mini-PROTEAN TGX Stain Free precast polyacrylamide 4–20% gels (2 citations) Precast 4–20% gradient polyacrylamide gels (2 citations)

4 protocols using precast 4 20 polyacrylamide gels

1

Protein Profiling via In-Gel Digestion

Check if the same lab product or an alternative is used in the 5 most similar protocols
For each protein sample (8‐h noninfected, 8‐h infected, 24‐h noninfected and 24‐h infected), 50 μg was loaded onto pre‐cast 4–20% polyacrylamide gels (Bio‐Rad) and samples were run two‐thirds of the way down the gel. Each lane was cut into eight equally sized pieces and each section cut into 1‐mm3 cubes. Gel pieces were washed with 100 mM NH4HCO3 and acetonitrile before reduction and alkylation with 10 mM DTT and 55 mM iodoacetamide, respectively. Proteins were subject to in‐gel digestion with sequencing‐grade trypsin (Roche) and digestions were carried out at 37°C overnight. The resulting peptides were cleaned using a C18 (POROS R2; Applied Biosystems, Waltham, MA, USA) column. Clean peptide samples were dried down to c. 10 μl using a vacuum centrifuge before being re‐adjusted to a final volume of 30 μl with 0.1% trifluoroacetic acid.
Howden A.J., Stam R., Martinez Heredia V., Motion G.B., ten Have S., Hodge K., Marques Monteiro Amaro T.M, & Huitema E. (2017). Quantitative analysis of the tomato nuclear proteome during Phytophthora capsici infection unveils regulators of immunity. The New Phytologist, 215(1), 309-322.
+ Open protocol
+ Expand
2

Western Blotting of Brain Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Input and pulldown gel samples were bath sonicated, boiled for 10 min, and separated by SDS-PAGE on precast 4-20% polyacrylamide gels (BioRad). Proteins were then transferred on PVDF membranes using a semi-dry transfer apparatus. For dot-blotting, each spot was added with 5 ug of brain lysate protein on a nitrocellulose membrane and allowed to air-dry for 1.5 hours before blocking. Membranes were blocked for 1 h at room temperature using OneBlock Western-FL Blocking Buffer (Genesee Scientific) after which the membranes were incubated with primary antibodies (Anti-O-GlcNAc RL2 1:3,000, Thermo MA1-072; Anti-Hsp27 1:3,000, Cell Signaling Technology #95357; Anti-αBC 1:3,000, Cell Signaling Technology #45584) in blocking buffer at 4 °C for 16 h. Membranes were washed with TBST (137 mM NaCl, 20 mM Tris, 0.1% Tween-20, pH 7.6, Cell Signaling Technology) 3 times for 10 min each, followed by incubation with HRP-conjugated anti-rabbit or anti-mouse secondary antibodies (1:10,000, Jackson ImmunoResearch 711-035-152 or 715-035-150) in blocking buffer. After 3x10 min washing in TBST, membranes were developed with Western ECL Substrates (Biorad) and imaged using a ChemiDoc XRS+ Imager (Bio-Rad).
Balana A.T., Levine P.M., Craven T.W., Mukherjee S., Pedowitz N.J., Moon S.P., Takahashi T.T., Becker C.F., Baker D, & Pratt M.R. (2021). O-GlcNAc modification of small heat shock proteins enhances their anti-amyloid chaperone activity. Nature chemistry, 13(5), 441-450.
+ Open protocol
+ Expand
3

Western Blotting of Brain Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Input and pulldown gel samples were bath sonicated, boiled for 10 min, and separated by SDS-PAGE on precast 4-20% polyacrylamide gels (BioRad). Proteins were then transferred on PVDF membranes using a semi-dry transfer apparatus. For dot-blotting, each spot was added with 5 ug of brain lysate protein on a nitrocellulose membrane and allowed to air-dry for 1.5 hours before blocking. Membranes were blocked for 1 h at room temperature using OneBlock Western-FL Blocking Buffer (Genesee Scientific) after which the membranes were incubated with primary antibodies (Anti-O-GlcNAc RL2 1:3,000, Thermo MA1-072; Anti-Hsp27 1:3,000, Cell Signaling Technology #95357; Anti-αBC 1:3,000, Cell Signaling Technology #45584) in blocking buffer at 4 °C for 16 h. Membranes were washed with TBST (137 mM NaCl, 20 mM Tris, 0.1% Tween-20, pH 7.6, Cell Signaling Technology) 3 times for 10 min each, followed by incubation with HRP-conjugated anti-rabbit or anti-mouse secondary antibodies (1:10,000, Jackson ImmunoResearch 711-035-152 or 715-035-150) in blocking buffer. After 3x10 min washing in TBST, membranes were developed with Western ECL Substrates (Biorad) and imaged using a ChemiDoc XRS+ Imager (Bio-Rad).
Balana A.T., Levine P.M., Craven T.W., Mukherjee S., Pedowitz N.J., Moon S.P., Takahashi T.T., Becker C.F., Baker D, & Pratt M.R. (2021). O-GlcNAc modification of small heat shock proteins enhances their anti-amyloid chaperone activity. Nature chemistry, 13(5), 441-450.
+ Open protocol
+ Expand
4

Western Blot Protein Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
Lysates containing equal amount of protein were run on pre-cast 4-20% polyacrylamide gels (Bio-Rad, Hercules, CA) followed by transfer onto Nitrocellulose blotting membrane (GE Healthcare, UK) using the wet-transfer method. The membranes were then blocked with PBS-T containing 5% milk, followed by overnight incubation with specific antibody in 1X PBS containing 5% milk. After multiple washing with 1X PBS-T, the membrane was incubated for 1 hr with HRP-conjugated secondary from the appropriate species. After multiple washes with 1X PBS-T, the membrane was exposed to SuperSignal® West Femto Maximum Sensitivity Substrate (ThermoScientific, Waltham, MA) per manufacturer’s instructions, followed by autoradiography.
Seth P., Hsieh P.N., Jamal S., Wang L., Gygi S.P., Jain M.K., Coller J, & Stamler J.S. (2019). Regulation of MicroRNA Machinery and Development by Interspecies S-Nitrosylation. Cell, 176(5), 1014-1025.e12.
+ 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!