Gradient 4 20 sds page gels

Manufactured by Bio-Rad

Sourced in United States

Gradient 4–20% SDS-PAGE gels are pre-cast polyacrylamide gels designed for the separation of proteins based on their molecular weight. The gels feature a linear gradient of polyacrylamide concentration from 4% to 20%, allowing for the effective separation of a wide range of protein sizes.

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Lab products found in correlation

Laemmli sample buffer, by Bio-Rad (2 mentions) Hsp90, by Enzo Life Sciences (1 mentions) Hsp70, by Enzo Life Sciences (1 mentions) Hsp40, by Cell Signaling Technology (1 mentions) Grp94, by Enzo Life Sciences (1 mentions) α tubulin, by Merck Group (1 mentions) Tonitrocellulose membranes, by Bio-Rad (1 mentions) Las4000 imager, by GE Healthcare (1 mentions) Pvdf membrane, by Cytiva (1 mentions) Horseradish peroxidase hrp conjugated secondary antibody, by Promega (1 mentions) Gapdh, by Merck Group (1 mentions) Complete antiprotease cocktail, by Roche (1 mentions) Las4000 image analyser, by GE Healthcare (1 mentions) Ripa buffer, by Thermo Fisher Scientific (1 mentions) Pvdf membrane, by GE Healthcare (1 mentions) Ecl detection reagent, by GE Healthcare (1 mentions) Dc protein assay, by Bio-Rad (1 mentions) Goat anti mouse igg horseradish peroxidase, by Merck Group (1 mentions) Fluorchem r system, by Bio-Techne (1 mentions) Goat anti rabbit igg horseradish peroxidase, by Merck Group (1 mentions)

Spelling variants of this product

SDS-PAGE (902 citations) SDS-PAGE gels (753 citations) 4–20% gradient SDS-PAGE gels (207 citations) 4–20% gradient SDS-PAGE (55 citations) 4–20% gradient SDS gel (6 citations) 4–15% SDS gradient gels (4 citations) SDS‐PAGE 4%–20% gel (3 citations) SDS-PAGE (4–20% gradient, (3 citations) PAGE gels (3 citations) 4–20% gradient PAGE gels (2 citations)

4 protocols using gradient 4 20 sds page gels

Platelet Proteome Analysis via Western Blot

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After treatment with the indicated inhibitors with or without
thrombin-mediated activation, platelet whole cell lysates were generated in TNES
buffer (50mM Tris pH7.5, 100mM NaCl, 2mM EDTA, 1% NP-40 supplemented with
protease and phosphatase inhibitors (cOmplete and PhosSTOP cocktail tablets,
respectively, Sigma-Aldrich, St. Louis, MO, USA). After cellular debris was
removed via high speed centrifugation, equal amounts of the lysates were
fractionated on gradient 4–20% SDS-PAGE gels and transferred to
nitrocellulose membranes (both from Bio-Rad, Hercules, CA, USA). The membranes
were analyzed for immunoreactivity to the following antibodies: Hsp40, (Cell
Signaling Technology, Danvers, MA, USA), Hsp70, Hsp90, and Grp94 (Enzo Life
Sciences, Farmingdale, NY, USA), and α-tubulin (EMD Millipore,
Burlington, MA, USA). Bound antibodies were detected via species-specific
horseradish peroxidase (HRP)- conjugated secondary antibodies at a 1:000
dilution (GE Healthcare, Pittsburgh, PA, USA), followed by reaction with Pico
ECL (Thermo Scientific, Rockford, IL, USA) to produce a chemiluminescent signal,
and exposed to x-ray film (Vita Scientific, College Park, MD, USA).

Jackson J.W., Rivera-Marquez G.M., Beebe K., Tran A.D., Trepel J.B., Gestwicki J.E., Blagg B.S., Ohkubo S, & Neckers L.M. (2020). Pharmacologic dissection of the overlapping impact of heat shock protein family members on platelet function. Journal of thrombosis and haemostasis : JTH, 18(5), 1197-1209.

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Western Blot Analysis of EGFR and α5 Integrin

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Cells were lysed in 1% Triton X-100, NaF [100 mmol/L], NaPPi [10 mmol/L], and Na₃VO₄ [1 mmol/L] in PBS, supplemented with complete anti-protease cocktail (Roche, Basel, Switzerland). A total of 10 µg of protein was separated on precast gradient 4–20% SDS-PAGE gels (Bio-Rad, Hercules, CA, USA) and transferred to polyvinylidene fluoride (PVDF) membranes (Amersham Bioscience, Buckinghamshire, UK). After blocking, membranes were probed with primary antibodies targeting EGFR (D38B1, #4267; Cell Signaling Technology, Danvers, MA, USA), α5 integrin (D7B7G, #98204S; Cell Signaling Technology), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Millipore, Molsheim, France). Immunological complexes were revealed with horseradish peroxidase (HRP)-conjugated secondary antibodies (Promega, Madison, WI, USA) at a 1/10,000 dilution. Revelation was performed with enhanced chemiluminescence (ECL; BioRad) using an LAS4000 imager (GE Healthcare, Dornstadt, Germany). GAPDH was used as housekeeping protein to serve as the loading control for all cell lysate samples. The quantification of non-saturated images was performed with ImageJ software. Analyses were performed on at least three independent experiments.

Cruz Da Silva E., Foppolo S., Lhermitte B., Ingremeau M., Justiniano H., Klein L., Chenard M.P., Vauchelles R., Abdallah B., Lehmann M., Etienne-Selloum N., Dontenwill M, & Choulier L. (2022). Bioimaging Nucleic-Acid Aptamers with Different Specificities in Human Glioblastoma Tissues Highlights Tumoral Heterogeneity. Pharmaceutics, 14(10), 1980.

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Protein Expression Analysis using Western Blot

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For protein expression analysis (Figure S1), after 3 washes in ice cold PBS, proteins were extract using RIPA buffer (Thermo Fisher Scientific, Braunsweig, Germany), according to manufacturer protocol, protein concentrations were evaluated using DC Protein assay (Bio-Rad, Hercules, CA, USA) and 20 µg of proteins were further analysed. For siRNA experiments, proteins were directly extracted in Laemmli sample buffer (Bio-Rad, Hercules, CA, USA). Proteins were separated on precast gradient 4–20% SDS-PAGE gels (Bio-Rad, Hercules, CA, USA) and transferred to a PVDF membrane (GE Healthcare, Dornstadt, Germany). Membranes were probed with primary antibodies: anti-EGFR antibody, anti-DNM2, anti-Rab5 and anti-LRP-1 at 1µg/mL and anti-GAPDH at 0.2 µg/mL in blocking solution (TBS—tween 0.1%, 5% non-fat dry milk). Immunological complexes were revealed with anti-rabbit or anti-mouse IgG coupled peroxidase antibodies using chemoluminescence (ECL detection reagent, GE Healthcare, Dornstadt, Germany) and visualised with a LAS4000 image analyser (GE Healthcare, Dornstadt, Germany). GAPDH was used as the loading control for all samples.

Cruz Da Silva E., Choulier L., Thevenard-Devy J., Schneider C., Carl P., Rondé P., Dedieu S, & Lehmann M. (2021). Role of Endocytosis Proteins in Gefitinib-Mediated EGFR Internalisation in Glioma Cells. Cells, 10(11), 3258.

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Western Blot Analysis of Parasitic Proteins

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Parasite extracts were resuspended in 2× Laemmli sample buffer (Bio-Rad) with 5% 2-mercaptoethanol (Sigma-Aldrich). Samples were boiled for 5 min at 95°C before separation on a gradient 4–20% SDS-PAGE gels (Bio-Rad). Samples were then transferred to nitrocellulose membrane using standard methods for semidry transfer (Bio-Rad). Membranes were probed with rabbit anti-HA (CS29F4 cat. no. 3724S, Cell Signaling Technology), mouse anti-c-Myc (9B11 cat. no. 2276, Cell Signaling Technology) or mouse anti-F₁B ATPase (made in house) all at a dilution of 1:5000, or rat anti-IMC10 (made in house) at a dilution 1:5000 overnight. Given the high molecular mass of ATPase-guanylyl cyclase, we used 4–20% Tris-acetate SDS gels (Invitrogen) as performed in Yang et al. (2019b) (link). Membranes were then washed and probed with either goat anti-mouse-IgG horseradish peroxidase or goat anti-rabbit-IgG horseradish peroxidase (Sigma-Aldrich) at a dilution of 1:10,000 for 1 h (GE Healthcare). Proteins were detected using SuperSignal West Femto substrate (Thermo Fisher) and imaged using the FluorChem R system (Biotechne). Full original western blots are shown in Fig. S6.

Oliveira Souza R.O., Jacobs K.N., Back P.S., Bradley P.J, & Arrizabalaga G. (2022). IMC10 and LMF1 mediate mitochondrial morphology through mitochondrion–pellicle contact sites in Toxoplasma gondii. Journal of Cell Science, 135(22), jcs260083.

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