Protein lysates of NPC cell pellets (Control, 45Q, and 81Q) were prepared using RIPA lysis buffer (RIPA Buffer, 1 mM PMSF, 5 μm Z-VAD, 1 mM NaVan, and 1 × Complete Protease Inhibitor Mixture tablets). Protein lysates were sonicated for 2 cycles of 30 s on, 30 s off. 35 μg of protein lysates were separated on 10% TGX FastCast gel (Biorad) and transferred on nitrocellulose membranes (Biorad). The following primary antibodies were used: CUL4A (1:1000; Proteintech Cat# 14851-1-AP, RRID:AB_2261175 ), H3 (1:2500, Abcam Cat# ab1791, RRID:AB_302613 ), H3K27me3 (1:500, Millipore Cat# 07-449, RRID:AB_310624 ), H3K9me1 (1:500; Thermo Fisher Scientific Cat# MA5-33385, RRID:AB_2815523 ), H3K9me2 (1:500; Thermo Fisher Scientific Cat# 720092, RRID:AB_2532802 ), and β-actin (1:5000; Sigma-Aldrich Cat# A5441, RRID:AB_476744 ) (Supplemental Table S1 ). Antibody validation proved by venders. Membranes were incubated with primary antibodies at 4 °C overnight. The following secondary antibodies (1:5000) were used: IRDye® 680RD Goat anti-Rabbit IgG and IRDye® 800RD Goat anti-Mouse IgG (LI-COR). The membrane was imaged using the LI-COR Imaging System and Odyssey V3.0 software (LI-COR), followed by intensity analysis with GelAnalyzer. Statistical significance was tested using a two tailed t-test with a P-value threshold of 0.05.
Cul4a
Manufactured by Proteintech
Sourced in China
CUL4A is a protein that is a component of the CUL4A-RING E3 ubiquitin ligase complex. This complex is involved in the regulation of various cellular processes through the ubiquitination of target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey v3, by LI COR (1 mentions)
Irdye 680rd goat anti rabbit igg, by LI COR (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
H3k27me3, by Merck Group (1 mentions)
β actin, by Merck Group (1 mentions)
Tgx fastcast gel, by Bio-Rad (1 mentions)
H3k9me1, by Thermo Fisher Scientific (1 mentions)
H3k9me2, by Thermo Fisher Scientific (1 mentions)
Irdye 800rd goat anti mouse igg, by LI COR (1 mentions)
P akt ser473, by Cell Signaling Technology (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
Cycloheximide chx solution, by Merck Group (1 mentions)
P akt thr308, by Cell Signaling Technology (1 mentions)
Siah2, by Proteintech (1 mentions)
Ubiquitin, by Proteintech (1 mentions)
β actin, by Proteintech (1 mentions)
Ship2, by Cell Signaling Technology (1 mentions)
Nedd4, by Proteintech (1 mentions)
Mg132, by Beyotime (1 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions)
3 protocols using cul4a
1
Epigenetic Changes in Huntington's Disease
2
Antibody Sources for Cell Signaling
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Antibodies against PHB2 (Cat#14085S), PHB1 (Cat#2426S), SHIP2 (Cat#2839S), Akt (Cat#9272S), pAkt-Ser473 (Cat#4060S), pAkt-Thr308 (Cat#13038S), p21 (Cat#2947S), p27 (Cat#3686S), Histone H3 (Cat#9715S), K63 (Cat#5621S), K48 (Cat#8081S), and Ki67 (Cat#34330SF) were purchased from Cell Signaling Technology (Beverly, MA, USA), antibodies against tGFP (Cat#TA150041) was purchased from OriGene (Rockville, MD, USA), antibodies against Flag (Cat#66008-3-Ig, Cat#20543-1-AP), HA (Cat#51064-2-AP, Cat#66006-2-Ig), His (Cat#66005-1-Ig), GST (Cat#66001-2-Ig), Ubiquitin (Cat#10201-2-AP), β-actin (Cat#66009-1-Ig), NEDD4 (Cat#21698-1-AP), SIAH2 (Cat#12651-1-AP), WWP2 (Cat#67274-1-Ig), Cul4A (Cat#14851-1-AP) and MUL1 (Cat#16133-1-AP) were purchased from Proteintech (Wuhan, China). MG132 (Cat#S1748) was purchased from Beyotime Biotechnology (Shanghai, China). Cycloheximide (CHX) solution was purchased from Sigma-Aldrich (St. Louis, MO, USA).
3
Antibody Panel for AMPK Signaling
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Primary antibodies against the following proteins were obtained from Cell Signaling Technology: p-AMPK (#2535, 1:1000), p-AMPK substrate motif (#5759, 1:1000), AMPKa (#2532, 1:2000), AMPKa2 (#2757, 1:2000), p-mTOR (#5536, 1:1000), mTOR (#2983, 1:2000), p-4EBP1 (#2855, 1:1000), p-S6K (#9205, 1:1000) and S6K (#9202, 1:2000); From Upstate: PROX1 (07-537, 1:2000); from Abcam: AMPKa1 (ab32047, 1:2000) and DDB1 (ab109027, 1:2000); from Sigma: Flag M2 (F3165, 1:5000) and β-Actin (A1978, 1:10000); from Thermo Fisher Scientific: HA (SG77) (71-5500, 1:3000); from Abclonal: p-S79 (customization, 1:1000), ACADM (A4567, 1:2000) and EHHADH (A13488, 1:2000); from Proteintech: CUL4A (14851-1-AP, 1:2000), CUL4B (12916-1-AP, 1:2000), BCKDHB (13685-1-AP, 1:2000) and ACADSB (13122-1-AP, 1:2000); from Cell Signaling Technology: anti-rabbit IgG, HRP-linked Antibody (#7074, 1:5000) and anti-mouse IgG, HRP-linked Antibody (#7076, 1:5000).
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