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 .
Phgdh hpa021241
Manufactured by Merck Group
PHGDH (HPA021241) is a recombinant protein that represents the Phosphoglycerate dehydrogenase enzyme. This enzyme is involved in the conversion of 3-phosphoglycerate to 3-phosphohydroxypyruvate as part of the serine biosynthesis pathway.
Automatically generated - may contain errors
Lab products found in correlation
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)
Iscript adv cdna synthesis kit, by Bio-Rad (2 mentions)
Psat1, by Novus Biologicals (2 mentions)
2 protocols using phgdh hpa021241
1
Protein and Gene Expression Analysis
2
Protein and Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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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 .
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