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Ab52818

Manufactured by Santa Cruz Biotechnology
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Sourced in United Kingdom

Ab52818 is a laboratory reagent designed for use in research applications. It is a primary antibody that can be used to detect and study specific target proteins or molecules in biological samples. The core function of Ab52818 is to bind to and identify the target of interest, facilitating further analysis and experimentation. No additional details or interpretations are provided.

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Lab products found in correlation

Bradford reagent, by Bio-Rad (2 mentions) Super signal chemiluminescence reagent, by Thermo Fisher Scientific (2 mentions) Immobilon p nitrocellulose membrane, by Merck Group (2 mentions) Bis tris gel, by Thermo Fisher Scientific (2 mentions) Ab128870, by BD (2 mentions) Western lightning ecl pro, by PerkinElmer (2 mentions) Hifi dna assembly master mix, by New England Biolabs (1 mentions) Transferrin, by Thermo Fisher Scientific (1 mentions)

3 protocols using ab52818

1

Western Blot Analysis of FASN KO Cells

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HAP1 WT and FASN KO cells were lysed in buffer F (10 mM Tris pH 7.05, 50 mM NaCl, 30 mM Na pyrophosphate, 50 mM NaF, 10% Glycerol, 0.5% Triton X-100) and centrifuged at 14,000 rpm for 10 minutes. The supernatant was collected and protein concentration was determined using Bradford reagent (BioRad). 10–30 μg protein was resolved on 4–12% Bis-Tris gels (Life Technologies) and transferred to Immobilon-P nitrocellulose membrane (Millipore) at 66V for 90 minutes. Subsequently, proteins were detected using anti-FASN (1:2,000, Abcam ab128870), anti-SREBP2 (1:250, BD Biosciences 557037), anti-LDLR (1:250, ab52818), anti-GAPDH (1:10,000, Santa Cruz 166574) and anti-β-Actin (1:10,000, Abcam ab8226) antibodies and proteins were visualized on X-ray film using Super Signal chemiluminescence reagent (Thermo Scientific) and Western Lightning ECL Pro (PerkinElmer). Scans of uncropped western blots are provided in Source Data Figure 5 and Source Data Extended Figures 1, 6.
Aregger M., Lawson K.A., Billmann M., Costanzo M., Tong A.H., Chan K., Rahman M., Brown K.R., Ross C., Usaj M., Nedyalkova L., Sizova O., Habsid A., Pawling J., Lin Z.Y., Abdouni H., Wong C.J., Weiss A., Mero P., Dennis J.W., Gingras A.C., Myers C.L., Andrews B.J., Boone C, & Moffat J. (2020). Systematic mapping of genetic interactions for de novo fatty acid synthesis identifies C12orf49 as a regulator of lipid metabolism. Nature metabolism, 2(6), 499-513.
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2

Western Blot Analysis of FASN KO Cells

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HAP1 WT and FASN KO cells were lysed in buffer F (10 mM Tris pH 7.05, 50 mM NaCl, 30 mM Na pyrophosphate, 50 mM NaF, 10% Glycerol, 0.5% Triton X-100) and centrifuged at 14,000 rpm for 10 minutes. The supernatant was collected and protein concentration was determined using Bradford reagent (BioRad). 10–30 μg protein was resolved on 4–12% Bis-Tris gels (Life Technologies) and transferred to Immobilon-P nitrocellulose membrane (Millipore) at 66V for 90 minutes. Subsequently, proteins were detected using anti-FASN (1:2,000, Abcam ab128870), anti-SREBP2 (1:250, BD Biosciences 557037), anti-LDLR (1:250, ab52818), anti-GAPDH (1:10,000, Santa Cruz 166574) and anti-β-Actin (1:10,000, Abcam ab8226) antibodies and proteins were visualized on X-ray film using Super Signal chemiluminescence reagent (Thermo Scientific) and Western Lightning ECL Pro (PerkinElmer). Scans of uncropped western blots are provided in Source Data Figure 5 and Source Data Extended Figures 1, 6.
Aregger M., Lawson K.A., Billmann M., Costanzo M., Tong A.H., Chan K., Rahman M., Brown K.R., Ross C., Usaj M., Nedyalkova L., Sizova O., Habsid A., Pawling J., Lin Z.Y., Abdouni H., Wong C.J., Weiss A., Mero P., Dennis J.W., Gingras A.C., Myers C.L., Andrews B.J., Boone C, & Moffat J. (2020). Systematic mapping of genetic interactions for de novo fatty acid synthesis identifies C12orf49 as a regulator of lipid metabolism. Nature metabolism, 2(6), 499-513.
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3

Characterizing LDLR Regulation by EXOC4 and EXOC8

Cited 1 time
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HuH7 cells were cultured in DMEM containing 10% FBS and penicillin/streptomycin. Cellular uptake assays were performed with fluorescent conjugates of LDL (Cayman Chemical, Ann Arbor MI, 10011229) or transferrin (ThermoFisher Scientific, Waltham MA, T35352). For immunoblotting, membranes were probed with antibodies against LDLR (Abcam, Cambridge UK, ab52818, 1:2000), β-actin (Santa Cruz Biotechnology, Dallas TX, sc-47778, 1:5000), EXOC4 (Abcam, ab205945, 1:1000), and EXOC8 (Santa Cruz, sc-515532, 1:500). For flow cytometry, cells were stained with a fluorescently-conjugated antibody against LDLR (R&D Systems, Minneapolois MN, FAB2148G). CRISPR-mediated gene disruption was performed by cloning gRNA sequences into BsmBI sites of pLentiCRISPRv2 (Addgene #52961, a gift from Feng Zhang[17 (link)]) or into BbsI sites of pX459 (Addgene #62988, a gift from Feng Zhang). Genotyping was performed by Sanger sequencing of PCR amplicons of genomic target sites with individual alleles deconvoluted by TIDE analysis of chromatograms[33 (link)]. Lentiviral expression constructs were generated by assembly of cDNA sequences (GE Healthcare Dharmacon, Lafayette CO, EXOC4 MHS6278-202759993 and EXOC8 MHS6278-202758964) and a blasticidin resistance cassette into the LeGO-ic2 plasmid (Addgene # 27345, a gift from Boris Fehse[34 (link)]) using HiFi DNA Assembly Master Mix (NEB, Ipswich MA).
Emmer B.T., Sherman E.J., Lascuna P.J., Graham S.E., Willer C.J, & Ginsburg D. (2021). Genome-scale CRISPR screening for modifiers of cellular LDL uptake. PLoS Genetics, 17(1), e1009285.
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