Sds gradient gels

Manufactured by Bio-Rad

4–15% SDS gradient gels are pre-cast polyacrylamide gels used for protein separation and analysis. They provide a linear separation range for proteins with molecular weights between 10 and 200 kDa. The gradient allows for efficient separation of proteins with a wide range of sizes in a single gel.

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Lab products found in correlation

Protein inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Anti e2f1, by Cell Signaling Technology (2 mentions) Anti rb, by Cell Signaling Technology (2 mentions) Anti foxa1, by Abcam (2 mentions) Anti tubulin, by Abcam (2 mentions) Anti v5, by Abcam (2 mentions) Anti e2f1, by Abcam (2 mentions) Anti lsd1, by Abcam (2 mentions) Anti corest, by Abcam (2 mentions) Anti h3, by Abcam (2 mentions) Nitrocellulose membrane, by Bio-Rad (2 mentions) Anti gapdh, by Abcam (2 mentions) Odyssey clx infrared imaging system, by LI COR (1 mentions) Pvdf fl membrane, by Merck Group (1 mentions) Survivin, by Cell Signaling Technology (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) C raf, by Cell Signaling Technology (1 mentions) 6 well plate, by Avantor (1 mentions) Pen strep, by Avantor (1 mentions) β actin, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

4–20% gradient SDS gel 4–15% Tris-HCl gradient SDS-PAGE gels Gradient 4–20% SDS-PAGE gels 4–20 gradient SDS acrylamide gels TGX Criterion gradient SDS polyacrylamide gels 4 to 20% gradient SDS-PAGE gels Gradient (4–15%) SDS-polyacrylamide gels Tris-Glycine SDS gradient gels Tris–glycine SDS-PAGE gradient (4–15% acrylamide) gels 10% SDS gel

4 protocols using sds gradient gels

Immunoprecipitation and Western Blotting

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For immunoprecipitation (IP), cells were lysed in Triton lysis buffer supplemented with protein inhibitor cocktails (Thermo Fisher Scientific), followed by a brief sonication, and then lysates were immunoprecipitated with anti-E2F1 (Abcam), anti-LSD1 (Abcam), or anti-CoREST (Abcam) antibodies. For immunoblotting, proteins were separated on 4–15% SDS gradient gels (Bio-Rad), transferred to nitrocellulose membranes (Bio-Rad), and then probed with primary antibodies, including anti-Rb (Cell Signaling), anti-tubulin (Abcam), anti-LSD1 (Abcam), anti-CoREST (Abcam), anti-E2F1 (Cell Signaling), anti-GAPDH (Abcam), anti-V5 (Abcam), anti-H3 (Abcam) and anti-FOXA1 (Abcam).

Han W., Liu M., Han D., Li M., Toure A.A., Wang Z., Besschetnova A., Patalano S., Macoska J.A., Gao S., Hansen He H, & Cai C. (2022). RB1 loss in castration-resistant prostate cancer confers vulnerability to LSD1 inhibition. Oncogene, 41(6), 852-864.

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Immunoprecipitation and Western Blotting

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SDS-PAGE and Western Blotting Quantification

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Protein samples were resolved on 4–20% SDS-gradient gels (BioRad, Hercules, CA), then transferred to PVDF-FL membrane (Millipore Burlington, MA). Detection and quantification was carried out using a LI-COR Odyssey infrared scanning system using fluorescently labelled secondary antibodies. All western blots were scanned using the Odyssey Clx infrared imaging system from LICOR.

D'Souza R.S., Lim J.Y., Turgut A., Servage K., Zhang J., Orth K., Sosale N.G., Lazzara M.J., Allegood J, & Casanova J.E. (2020). Calcium-stimulated disassembly of focal adhesions mediated by an ORP3/IQSec1 complex. eLife, 9, e54113.

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GDA and Compound 6a Effects on HCT-116 Cells

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HCT-116 cells were grown in DMEM (Corning, 10–027-CV) containing 1% Pen-Strep (VWR, K952–100ML) and 10% FBS+ (Atlas Biologicals, F-0500-D), and plated in a 6-well plate (VWR, 10861–696). At ∼80%, cells were treated with GDA at 500 nM and compound 6a at 100 nM, 250 nM, 1.25 μM and 2.5 μM for 24 hours.
Cell culture samples were lysed using RIPA buffer containing protease and phosphatase inhibitors. Lysate protein concentrations were determined by BCA assay (Thermo). Samples were separated by SDS-PAGE using 4–20% SDS gradient gels (BioRad). Gel contents were transferred to PVDF membranes. Membranes were blocked with 7% non-fat dry milk solution. Blots were probed with antibodies raised against Erk5 (Cell Signaling, 3552S), CDK4, (Cell Signaling, 12790S), CDK6 (Cell Signaling, 3136S), beta-Actin (Cell Signaling, 3700S), Survivin (Cell Signaling, 2808S), c-Raf (Cell Signaling, 7065S). Antibody dilutions were 1:1000 unless otherwise stated, and all secondary antibodies were used at 1:1000 (Southern Biotech). Blots were developed using ECL (Amersham, 45–000-999) on a ChemiDoc Imaging System (BioRad). Densitometry was performed using Image Lab software (BioRad).

Mishra S.J., Liu W., Beebe K., Banerjee M., Kent C.N., Munthali V., Koren J., Taylor J.A., Neckers L.M., Holzbeierlein J, & Blagg B.S. (2021). The Development of Hsp90β-selective Inhibitors to Overcome Detriments Associated with pan-Hsp90 Inhibition. Journal of medicinal chemistry, 64(3), 1545-1557.

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