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Tris glycine sds gels

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4–20% Tris Glycine SDS gels are pre-cast polyacrylamide gel electrophoresis (PAGE) gels used for the separation of proteins based on their molecular weight. These gels have a gradient of 4% to 20% polyacrylamide concentration, which allows for the effective separation of a wide range of protein sizes. The gels are made with Tris-Glycine-SDS buffer system, which is a standard buffer system for SDS-PAGE.

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Lab products found in correlation

Horseradish peroxidase conjugated secondary antibody, by Cell Signaling Technology (2 mentions) Supersignal west pico, by Thermo Fisher Scientific (2 mentions) Polyvinylidene difluoride membrane, by Bio-Rad (2 mentions) Ripa buffer, by Cell Signaling Technology (2 mentions) Shmt2, by Cell Signaling Technology (2 mentions) Shmt1, by Cell Signaling Technology (2 mentions) Mthfd1, by Proteintech (2 mentions) Phenylmethylsulphonyl fluoride, by Merck Group (2 mentions) Protease and phosphatase inhibitor cocktail, by Merck Group (2 mentions) Bca protein assay, by Thermo Fisher Scientific (2 mentions) Pvdf membrane, by Merck Group (2 mentions) Irdye800, by Thermo Fisher Scientific (1 mentions) Odyssey infrared imager, by LI COR (1 mentions) Hmgb1, by Thermo Fisher Scientific (1 mentions) Alexa fluor 680, by Thermo Fisher Scientific (1 mentions) Glyceraldehyde 3 phosphate dehydrogenase, by Cell Signaling Technology (1 mentions) Mouse α flag clone m2, by Merck Group (1 mentions) Goat α rabbit hrp, by Bio-Rad (1 mentions) Rabbit α gapdh, by Santa Cruz Biotechnology (1 mentions) Goat α mouse hrp, by Bio-Rad (1 mentions)

Spelling variants of this product

4–20% Tris-glycine gels (192 citations) Glycine (46 citations) 10% SDS gel (31 citations) Tris/Glycine/SDS (15 citations) Tris gels (3 citations) Tris-glycine 4–20% SDS-PAGE gels (3 citations) Tris-glycine gels with Tris/glycine/SDS buffer (3 citations) 10% Tris–glycine precast SDS-PAGE gels (3 citations) Tris-Glycine SDS gradient gels (2 citations) Tris-Glycine-eXtendet SDS-PAGE gels (2 citations)

5 protocols using tris glycine sds gels

1

Western Blot Analysis of One-Carbon Metabolism Enzymes

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Tissue lysates were prepared in Kontes tubes with disposable pestles using RIPA Buffer (Cell Signaling Technology) supplemented with phenylmethylsulphonyl fluoride (Sigma), and protease and phosphatase inhibitor cocktails (Roche). The BCA protein assay (Thermo) was used to quantify protein concentrations. Equal amounts of protein (15–30 μg) were separated on 4–20% Tris Glycine SDS gels (BioRad) and transferred to polyvinylidene difluoride membranes (BioRad). Membranes were blocked for 30 minutes at room temperature with 5% milk in TBS supplemented with 0.1% Tween20 (TBST) then incubated with primary antibodies overnight at 4°C. After incubating with horseradish peroxidase conjugated secondary antibodies (Cell Signaling Technology), membranes were developed using SuperSignal West Pico or Femto chemiluminescence reagents (Thermo). Blots were stripped with 1% SDS, 25mM glycine (pH 2) before re-probing. The following primary antibodies were used for western blot analyses:ALDH1L2 (LifeSpan Bio; LS-C178510), DHFR (LifeSpan Bio; LS-C138829), MTHFR (LifeSpan Bio; LS-C157974), SHMT1 (Cell Signaling; 12612S), SHMT2 (Cell Signaling; 12762S), MTHFD1 (ProteinTech; 10794-1-AP), MTHFD2 (ProtenTech; 12270-1-AP), Actin (Abcam, ab8227).
Piskounova E., Agathocleous M., Murphy M.M., Hu Z., Huddlestun S.E., Zhao Z., Leitch A.M., Johnson T.M., DeBerardinis R.J, & Morrison S.J. (2015). Oxidative stress inhibits distant metastasis by human melanoma cells. Nature, 527(7577), 186-191.
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2

Western Blot Analysis of HMGB1 and TLR4

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Thirty micrograms of renal cortex was homogenized, and the whole homogenate was used for Western blot analysis as previously described [27 (link)]. Briefly, samples were resolved on 4–20% Tris-glycine-SDS gels (Bio-Rad, Hercules, CA) and proteins were transferred to PVDF membranes (Millipore Sigma, Burlington, MA). Protein expression was determined using two-color immunoblots using primary antibodies to HMGB1 (cat# MA1–20338, 1:1000 dilution; Thermo Fisher Scientific, Lafayette, CO), and TLR4 (cat# 217274, 1:300 dilution; Abcam, Cambridge, MA). Specific protein bands were detected using an Odyssey infrared imager (LI-COR Biosciences, Lincoln, NE) with AlexaFluor 680 and IRDye800 (Molecular Probes, Eugene, OR) conjugated secondary antibodies. Protein concentrations were determined by standard BCA reagent (Thermo Scientific) using BSA as the standard. Protein loading was normalized to glyceraldehyde 3-phosphate dehydrogenase (Cell Signaling Technologies, Danvers, Massachusetts).
Mohamed R., Rafikova O., O’Connor P.M, & Sullivan J.C. (2020). Greater high-mobility group box 1 in male compared with female spontaneously hypertensive rats worsens renal ischemia–reperfusion injury. Clinical science (London, England : 1979), 134(13), 1751-1762.
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3

Western Blot Analysis of One-Carbon Metabolism Enzymes

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Tissue lysates were prepared in Kontes tubes with disposable pestles using RIPA Buffer (Cell Signaling Technology) supplemented with phenylmethylsulphonyl fluoride (Sigma), and protease and phosphatase inhibitor cocktails (Roche). The BCA protein assay (Thermo) was used to quantify protein concentrations. Equal amounts of protein (15–30 μg) were separated on 4–20% Tris Glycine SDS gels (BioRad) and transferred to polyvinylidene difluoride membranes (BioRad). Membranes were blocked for 30 minutes at room temperature with 5% milk in TBS supplemented with 0.1% Tween20 (TBST) then incubated with primary antibodies overnight at 4°C. After incubating with horseradish peroxidase conjugated secondary antibodies (Cell Signaling Technology), membranes were developed using SuperSignal West Pico or Femto chemiluminescence reagents (Thermo). Blots were stripped with 1% SDS, 25mM glycine (pH 2) before re-probing. The following primary antibodies were used for western blot analyses:ALDH1L2 (LifeSpan Bio; LS-C178510), DHFR (LifeSpan Bio; LS-C138829), MTHFR (LifeSpan Bio; LS-C157974), SHMT1 (Cell Signaling; 12612S), SHMT2 (Cell Signaling; 12762S), MTHFD1 (ProteinTech; 10794-1-AP), MTHFD2 (ProtenTech; 12270-1-AP), Actin (Abcam, ab8227).
Piskounova E., Agathocleous M., Murphy M.M., Hu Z., Huddlestun S.E., Zhao Z., Leitch A.M., Johnson T.M., DeBerardinis R.J, & Morrison S.J. (2015). Oxidative stress inhibits distant metastasis by human melanoma cells. Nature, 527(7577), 186-191.
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4

Western Blot Antibody Characterization

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The antibodies used in these assays comprised monoclonal mouse anti(α)-influenza A NP blend clone A1/A3 (Millipore, 1:1,000 dilution); mouse α-Flag clone M2 (Sigma, 1:5,000); mouse α-HPV16 E7 clone ED17 (Santa Cruz, 1:200); mouse α-HPV16 E7 clone 8C9 (Invitrogen, 1:150); mouse α-HPV16 E6 clone 6F4 (Euromedex, 1:200); mouse α-HPV16 L1 Camvir-1 (BD Biosciences, 1:5,000); mouse α-pRb (1:500 dilution); polyclonal rabbit α-HPV16 L2 11–200 (described in [42 (link)], 1:5,000) and rabbit α-GAPDH (Santa Cruz, 1:1.000); goat α-rabbit HRP and goat α-mouse HRP (both from Biorad,1:25,000). Proteins were separated on 10 to 15% gradient Tris-glycine SDS gels (Biorad) using the Tris-glycine SDS buffer system. Western blotting was performed by electrophoretic transfer of proteins to a polyvinyl difluoride (PVDF) membrane (Millipore).
Jindra C., Huber B., Shafti-Keramat S., Wolschek M., Ferko B., Muster T., Brandt S, & Kirnbauer R. (2015). Attenuated Recombinant Influenza A Virus Expressing HPV16 E6 and E7 as a Novel Therapeutic Vaccine Approach. PLoS ONE, 10(9), e0138722.
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5

Comprehensive Immunoblotting Analysis of Signaling

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Antibodies against Vinculin (13901), AKT1 (2938), AKT2 (3063), pAKT T308 (13038), pAKT S473 (4060), c-PARP (D214) (5625), pPRAS40 T246 (13175), PRAS40 (8858), pAKT1 S473 (9018), pAKT2 S474 (8599), panAKT (4685), pBAD S136 (4366), BAD (9239), pChk2 T68 (2197), Chk2 (6334), LC3B (3868), GAPDH (5174), pS6 S235/236 (4858) and S6 (2217) were purchased from Cell Signalling. Cell lysates were prepared using 1× lysis buffer (9803, Cell Signalling) supplemented with 1% sodium dodecyl sulfate (SDS) (Sigma, L3771). Following sonication, lysates were quantified using the DC assay (Bio-Rad, 5000111) and equal amount of protein lysates were loaded into Tris-Glycine-SDS gels (Bio-Rad, 567-1095). Proteins were transferred into nitrocellulose membrane (Bio-Rad, 1704271) using the Trans-Blot Turbo Transfer System (Bio-Rad, 1704150) and were detected using Enhanced Chemiluminescence (ECL, Amersham, RPN2106/Bio-Rad, 1705062). Signal was captured on film (Amersham Hyper films ECL, 28906836) and developed using the Xorgaph X4 imaging system. Western blots were performed at least twice and a representative experiment is shown.
Kostaras E., Kaserer T., Lazaro G., Heuss S.F., Hussain A., Casado P., Hayes A., Yandim C., Palaskas N., Yu Y., Schwartz B., Raynaud F., Chung Y.L., Cutillas P.R, & Vivanco I. (2020). A systematic molecular and pharmacologic evaluation of AKT inhibitors reveals new insight into their biological activity. British Journal of Cancer, 123(4), 542-555.
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