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Psat1

Manufactured by Novus Biologicals
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PSAT1 is a lab equipment product offered by Novus Biologicals. It is a tool designed for laboratory use, but the specific core function of the product is not available in a detailed, unbiased, and factual description without additional interpretation or extrapolation.

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Lab products found in correlation

Phgdh hpa021241, by Merck Group (2 mentions) Iscript adv cdna synthesis kit, by Bio-Rad (2 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions) Proteinase inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Trizol, by Thermo Fisher Scientific (2 mentions) β actin, by Merck Group (2 mentions) Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (1 mentions) α actin, by Abcam (1 mentions) Phgdh, by Merck Group (1 mentions) Immobilon fl polyvinylidene fluoride pvdf membranes, by Merck Group (1 mentions) Odyssey image analysis system, by LI COR (1 mentions) α tubulin, by Merck Group (1 mentions) Alexa fluor 680, by Thermo Fisher Scientific (1 mentions) Irdye 800, by Rockland Immunochemicals (1 mentions) Parp1, by Cell Signaling Technology (1 mentions) Phospho t389 s6 kinase, by Cell Signaling Technology (1 mentions) Phospho ampkα t172, by Cell Signaling Technology (1 mentions) Phospho ulk1 s757, by Cell Signaling Technology (1 mentions) S6 ribosomal protein, by Cell Signaling Technology (1 mentions) β actin, by Cell Signaling Technology (1 mentions)

4 protocols using psat1

1

Protein and Gene Expression Analysis

Cited 1 time
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These were carried out as described previously (Wang et al., 2020 (link); Yan et al., 2005 (link)). For western blotting, cells were lysed in RIPA buffer (50 mM Tris-HCl, pH7.4, 150 mM NaCl, 1% NP-40. 0.1% SDS, 0.5% sodium deoxycholate, 1 mM EDTA) supplemented with proteinase inhibitor cocktails (PI78430, Fisher Scientific). The following antibodies were used: ATF3 (sc-188X, 1:10,000), ATF4 (sc-200, 1:1,000), p300 (sc-584, 1:200), and p53 (sc-126, 1:1000) from Santa Cruz; PHGDH (HPA021241, 1:300), β-actin (A2228, 1:10,000) from Sigma-Aldrich; and PSAT1 (21020002, 1:3,000) from Novus. For qRT-PCR, total RNA was isolated using TRIzol (Invitrogen), following by reverse transcription using 1 μg of total RNA and the iScript Adv cDNA synthesis kit (Bio-Rad). cDNAs were subjected to real-time PCR using SYBR Green (Bimaker) and the StepOnePlus Real-time PCR System (Applied Biosystems). The primers are listed in Table S4.
Li X., Gracilla D., Cai L., Zhang M., Yu X., Chen X., Zhang J., Long X., Ding H.F, & Yan C. (2021). ATF3 promotes the serine synthesis pathway and tumor growth under dietary serine restriction. Cell reports, 36(12), 109706.
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2

Western Blot Analysis of Cellular Proteins

Cited 1 time
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Whole cell extracts for Western blotting were prepared as described previously (16 (link)). Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto Immobilon-FL Polyvinylidene fluoride (PVDF) membranes (Millipore) according to standard procedures (Novex). Blots were probed with following antibodies: XRCC1 (Neomarkers, MS-1393-P0), DNA polymerase β (raised in-house and affinity-purified), α-tubulin (Sigma, T6199), MCM4 (abcam, ab4459-50), RP-A p32 (Bethyl, A300-244A), PCNA (Santa Cruz, sc-56), α-actin (Abcam, ab6276), PARP-1 (raised in-house and affinity-purified), p21 (Cell signaling, 12D1), PSAT1 (Novusbio, 21020002), PHGDH (Sigma, HPA021241), p53 (Santa Cruz, sc-126), PNKP (Abnova, H00011284-B01). Secondary antibodies conjugated with Alexa Fluor 680 (Molecular Probes) and IRDye® 800 (Rockland) fluorescent dyes were used. Detection and quantification was carried out using an Odyssey image analysis system (Li-Cor Biosciences).
Markkanen E., Fischer R., Ledentcova M., Kessler B.M, & Dianov G.L. (2015). Cells deficient in base-excision repair reveal cancer hallmarks originating from adjustments to genetic instability. Nucleic Acids Research, 43(7), 3667-3679.
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3

Protein and Gene Expression Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
These were carried out as described previously (Wang et al., 2020 (link); Yan et al., 2005 (link)). For western blotting, cells were lysed in RIPA buffer (50 mM Tris-HCl, pH7.4, 150 mM NaCl, 1% NP-40. 0.1% SDS, 0.5% sodium deoxycholate, 1 mM EDTA) supplemented with proteinase inhibitor cocktails (PI78430, Fisher Scientific). The following antibodies were used: ATF3 (sc-188X, 1:10,000), ATF4 (sc-200, 1:1,000), p300 (sc-584, 1:200), and p53 (sc-126, 1:1000) from Santa Cruz; PHGDH (HPA021241, 1:300), β-actin (A2228, 1:10,000) from Sigma-Aldrich; and PSAT1 (21020002, 1:3,000) from Novus. For qRT-PCR, total RNA was isolated using TRIzol (Invitrogen), following by reverse transcription using 1 μg of total RNA and the iScript Adv cDNA synthesis kit (Bio-Rad). cDNAs were subjected to real-time PCR using SYBR Green (Bimaker) and the StepOnePlus Real-time PCR System (Applied Biosystems). The primers are listed in Table S4.
Li X., Gracilla D., Cai L., Zhang M., Yu X., Chen X., Zhang J., Long X., Ding H.F, & Yan C. (2021). ATF3 promotes the serine synthesis pathway and tumor growth under dietary serine restriction. Cell reports, 36(12), 109706.
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4

Western Blot Analysis of Cellular Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in buffer containing 50 mM Tris pH 7.4, 1% Nonidet P-40, 0.25% sodium deoxycholate, 1 mM EDTA, 150 mM NaCl, 1 mM dithiothreitol, 1 mM sodium orthovanadate, 20 mM sodium fluoride, 10 mM beta-glycerophosphate, 10 mM sodium pyrophosphate, 2 ug ml−1 aprotinin, 2 ug ml−1 leupeptin and 0.7 ug ml−1 pepstatin. Western blot analysis was performed using standard protocols, and the following commercial antibodies were used as probes: ASNS (Proteintech 14681-1-AP, 1:1000), ATF4 (Cell Signaling 11815, 1:500), phospho-T389 S6 kinase (Cell Signaling 9234, 1:500), S6 kinase (Cell Signaling 2708, 1:1000), phospho-S235/235 S6 ribosomal protein (Cell Signaling 4858, 1:3000), S6 ribosomal protein (Cell Signaling 2217, 1:1000), phospho-S1859 CAD (Cell Signaling 70307, 1:500), CAD (Cell Signaling 11933, 1:1000), 4E-BP1 (Cell Signaling 9644, 1:500), phospho-S757 ULK1 (Cell Signaling 14202, 1:1000), ULK1 (Cell Signaling 8054, 1:1000), phospho-T172 AMPKα (Cell Signaling 2535, 1:500), AMPKα (Cell Signaling 2532, 1:1000), PHGDH (Cell Signaling 13428, 1:500), β-Actin (Cell Signaling 4970, 1:1000), PSAT1 (Novus 89-004-606, 1:500), and a-tubulin (Sigma T6074, 1:10,000).
Krall A.S., Mullen P.J., Surjono F., Momcilovic M., Schmid E.W., Halbrook C.J., Thambundit A., Mittelman S.D., Lyssiotis C.A., Shackelford D.B., Knott S.R, & Christofk H.R. (2021). Asparagine couples mitochondrial respiration to ATF4 activity and tumor growth. Cell metabolism, 33(5), 1013-1026.e6.
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