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ABCG1 protein, human

ABCG1 protein is a member of the ATP-binding cassette (ABC) transporter superfamily.
It plays a key role in cellular cholesterol homeostasis and is involved in the regulation of cholesterol efflux from cells.
ABCG1 is expressed in various tissues, including macrophages, and its dysregulation has been implicated in the pathogenesis of cardiovascular and metabolic disorders.
Researchers studying ABCG1 can leverage the power of PubCompare.ai, an AI-driven tool that helps identify the most reproducible and effective research protocols from the literature, preprints, and patents.
This streamlines the research process and takes ABCG1 protein studies to new heights by optimizing the selection of experimental methods.

Most cited protocols related to «ABCG1 protein, human»

1
Quantification of Cholesterol Efflux Capacity
Cholesterol efflux capacity was quantified in blood samples from the cohort of healthy volunteers as described previously.17 (link) This assay quantifies total efflux mediated by pathways of known relevance in cholesterol efflux from macrophages (i.e., ATP-binding cassette transporter A1 [ABCA1] and G1 [ABCG1], scavenger receptor B1, and aqueous diffusion).17 (link) Each sample was run in triplicate, with a mean coefficient of variation of 4.3%. Values were normalized by dividing the efflux capacity of individual patients by the efflux capacity of a serum pool run with each assay.
Cholesterol efflux capacity in the coronary disease and pharmacologic-study cohorts was quantified with the use of a slightly modified method designed to increase throughput. J774 cells, derived from a murine macrophage cell line, were plated and radiolabeled with 2 μCi of 3H-cholesterol per milliliter. ABCA1 was up-regulated by means of a 6-hour incubation with 0.3 mM 8-(4-chlorophenylthio)-cyclic AMP. Subsequently, efflux mediums containing 2.8% apolipoprotein B–depleted serum were added for 4 hours. All steps were performed in the presence of the acyl–coenzyme A:cholesterol acyltransferase inhibitor CP113,818 (2 μg per milliliter). In a pilot study involving serum samples from 20 healthy volunteers, results from the original assay procedure17 (link) and the modified method were strongly correlated (r = 0.85).
Liquid scintillation counting was used to quantify the efflux of radioactive cholesterol from the cells. The quantity of radioactive cholesterol incorporated into cellular lipids was calculated by means of isopropanol extraction of control wells not exposed to patient serum. Percent efflux was calculated by the following formula: [(microcuries of 3H-cholesterol in mediums containing 2.8% apolipoprotein B–depleted serum – microcuries of 3H-cholesterol in serum-free mediums) ÷ microcuries of 3H-cholesterol in cells extracted before the efflux step] × 100. All assays were performed in duplicate. To correct for interassay variation across plates, a pooled serum control from five healthy volunteers was included on each plate, and values for serum samples from patients were normalized to this pooled value in subsequent analyses. Additional studies that were performed to validate the measurement of cholesterol efflux capacity are described in the Supplementary Appendix.
Khera A.V., Cuchel M., de la Llera-Moya M., Rodrigues A., Burke M.F., Jafri K., French B.C., Phillips J.A., Mucksavage M.L., Wilensky R.L., Mohler E.R., Rothblat G.H, & Rader D.J. (2011). Cholesterol Efflux Capacity, High-Density Lipoprotein Function, and Atherosclerosis. The New England journal of medicine, 364(2), 127-135.
Publication 2011
ABCA1 protein, human ABCG1 protein, human Acyl Coenzyme A Apolipoproteins B Biological Assay BLOOD Cell Lines Cells Cholesterol Culture Media Cyclic AMP Diffusion Healthy Volunteers Heart Disease, Coronary Isopropyl Alcohol Lipids Macrophage Mus Patients Radioactivity Scavenger Receptor Serum Sterol O-Acyltransferase
2
Generation and Culture of Murine Bone Marrow-Derived Macrophages
Cryopyrin−/−, caspase-1−/−, Abca1−/−, Abcg1−/−, Abca1−/−Abcg1−/−, Kir6.1−/−, Kir6.2−/−, and SUR2−/− mice have been described previously (Miki et al., 1998 (link), 2002 (link); Schott et al., 2004 (link); Mariathasan et al., 2006 (link); Stoller et al., 2007 (link); Yvan-Charvet et al., 2008 (link)). P2X7−/− mice were obtained from Lexicon Pharmaceuticals. Mice were housed in a pathogen-free facility. All experiments were conducted in compliance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee at Genentech. BMDMs were prepared as described previously (Lamkanfi et al., 2007 (link)). In brief, BMDMs were isolated from femurs of 6–12-wk-old mice and were cultured in Iscove's modified Dulbecco's medium containing 10% heat-inactivated FBS, 20% L cell–conditioned medium, 100 U/ml penicillin, and 100 mg/ml streptomycin at 37°C in a humidified atmosphere containing 5% CO2. After 5–7 d of incubation, cells were collected and plated in 6- or 24-well plates in Iscove's modified Dulbecco's medium containing 10% heat-inactivated FBS and 100 mg/ml thymidine and antibiotics. Macrophages were cultured for an additional 24 h before use.
Lamkanfi M., Mueller J.L., Vitari A.C., Misaghi S., Fedorova A., Deshayes K., Lee W.P., Hoffman H.M, & Dixit V.M. (2009). Glyburide inhibits the Cryopyrin/Nalp3 inflammasome. The Journal of Cell Biology, 187(1), 61-70.
Publication 2009
ABCA1 protein, human ABCG1 protein, human Animals, Laboratory Antibiotics Atmosphere Caspase 1 Cells Culture Media, Conditioned Femur Institutional Animal Care and Use Committees L Cells Macrophage Mus NLRP3 protein, human Pathogenicity Penicillins Pharmaceutical Preparations Streptomycin Thymidine
3
Macrophage Isolation and Manipulation
Primary peritoneal macrophages were obtained from thioglycollate-treated mice 4 days after injection. For bone marrow-derived macrophages, bone marrow was isolated from femurs and tibias, and differentiated in DMEM supplemented with 20% fetal bovine serum (FBS), 30% L929 conditioned medium and antibiotics for 6–7 days. MEFs were immortalized by the SV40 Large T antigen retrovirus and selected with puromycin. Immortalized bone marrow-derived macrophages were obtained as previously described (Blasi et al., 1985 (link); Gandino and Varesio, 1990 (link)). To reconstitute immortalized bone marrow derived macrophages from LXR-deficient mice with LXR wild-type or mutants, retrovirus supernatants from Phoenix E cells transfected with the expression vectors were infected to immortalized macrophages. The iBMDM stably expressing wild-type LXRα, LXRβ or mutants were obtained by 300 µg/ml of hygromycin B (Invitrogen). For RNAi experiments in macrophages, cells were transfected with control or ON-TARGETplus SMARTpool siRNAs (25 nM, Dharmacon) targeted RXRα, RXRβ, Ubc9, Hdac4, ABCA1, ABCG1 and TRAF6 using Darmafect 4 (Dharmacon). Cells were used for experiments after 48 hr incubation and target gene knockdown was validated by real-time PCR and Western Blot. For macrophage inflammatory responses, cells were placed in DMEM containing 0.5% FBS, 5 μM simvastatin, 100 μM mevalonic acid and 1 μM GW3965 or DMSO overnight. Cells were then stimulated with 10 ng/ml LPS or vehicle. For cellular cholesterol modification, bone marrow-derived macrophages were placed in DMEM containing 0.5% FBS, 5 μM simvastatin plus 100 μM mevalonic acid for 4 hr, then incubated with complexes of randomly methylated cyclodextrin (Trappsol) and cholesterol (Sigma–Aldrich) (CD/Chol, 100 μM) to overload with cholesterol (Klein et al., 1995 (link)) or hydroxypropyl-β-cyclodextrin (Trappsol) (CD, 10 mM) to deplete cholesterol for 1 hr and stimulated with LPS (10 ng/ml) for 4 hr.
Ito A., Hong C., Rong X., Zhu X., Tarling E.J., Hedde P.N., Gratton E., Parks J, & Tontonoz P. (2015). LXRs link metabolism to inflammation through Abca1-dependent regulation of membrane composition and TLR signaling. eLife, 4, e08009.
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Publication 2015
ABCA1 protein, human ABCG1 protein, human Antibiotics Bone Marrow Cells Cholesterol Cloning Vectors Culture Media, Conditioned Cyclodextrins Femur Fetal Bovine Serum Gene Knockdown Techniques GW 3965 Hygromycin B Hypromellose Inflammation Large T-Antigen Macrophage Macrophages, Peritoneal Mevalonic Acid Mus Puromycin Real-Time Polymerase Chain Reaction Retroviridae RNA, Small Interfering RNA Interference Simian virus 40 Simvastatin simvastatin acid Sulfoxide, Dimethyl Thioglycolates Tibia TNF Receptor Associated Factor 6 Western Blot
4
Comparative Analysis of ABCG Transporters
The wild-type Arabidopsis background Col-0 (CS60000) and the T-DNA insertions of AtABCG1 (abcg1-1: SALK_061511; abcg1-2: SALK_055389) and AtABCG16 (abcg16-1: SALK_087501; abcg16-2: SALK_119868C) were obtained from Arabidopsis Biological Resource Center (ABRC)8. Pineapple (Ananas comosus) variety MD2 was collected by Qin Lab9. Pineapple were grown in plastic pots with soil mix [peat moss: perlite = 2:1 (v/v)] and placed in greenhouse at about 30°C with light availability of 60–70 mMolL-1 photons m-2s-1 under 70% humidity with 16 h light/8 h dark photoperiod. Arabidopsis was grown with the conditions as described by Cai et al. (2017) (link).
Chen P., Li Y., Zhao L., Hou Z., Yan M., Hu B., Liu Y., Azam S.M., Zhang Z., Rahman Z.U., Liu L, & Qin Y. (2017). Genome-Wide Identification and Expression Profiling of ATP-Binding Cassette (ABC) Transporter Gene Family in Pineapple (Ananas comosus (L.) Merr.) Reveal the Role of AcABCG38 in Pollen Development. Frontiers in Plant Science, 8, 2150.
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Publication 2017
ABCG1 protein, human Ananas Arabidopsis Biopharmaceuticals Humidity Insertion Mutation Light Marijuana Abuse Perlite Pineapple Sphagnopsida
5
Macrophage RNA Expression Profiling
Total RNA was extracted from peritoneal macrophages using a commercially available kit (QIAGEN; Hilden, Germany). The gene expressions were assessed by real-time RT-PCR using the TaqMan gene expression assay and a sequence detection system (ABI PRISM 7900, Life Technologies, Carlsbad, CA, USA), which used cDNA synthesized from isolated RNA samples to detect the expression of target genes, such as Lox-1; Mm00454586_m1, CD36; Mm01135198_ml, ACAT1; Mm00507463_ml, ABCA1; Mm00442646_ml, and ABCG1; Mm00437390_m1. TaqMan gene expression assays for mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was purchased from Life Technologies. The levels of these gene expressions were normalized using GAPDH as an internal control.
Terasaki M., Hiromura M., Mori Y., Kohashi K., Nagashima M., Kushima H., Watanabe T, & Hirano T. (2015). Amelioration of Hyperglycemia with a Sodium-Glucose Cotransporter 2 Inhibitor Prevents Macrophage-Driven Atherosclerosis through Macrophage Foam Cell Formation Suppression in Type 1 and Type 2 Diabetic Mice. PLoS ONE, 10(11), e0143396.
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Publication 2015
ABCA1 protein, human ABCG1 protein, human Biological Assay DNA, Complementary Gene Expression Glyceraldehyde-3-Phosphate Dehydrogenases Macrophages, Peritoneal Mus prisma Real-Time Polymerase Chain Reaction

Most recents protocols related to «ABCG1 protein, human»

1
Quantifying Membrane Transporter Proteins
Cells were washed with phosphate-buffered saline (PBS) and lysed in lysis buffer (50 mM Tris-Cl (pH 7.5), 150 mM NaCl, and 1% Triton X-100) containing 100 μg/mL 4-amidinophenylmethanesulfonyl fluoride, 2 μg/mL leupeptin, and 2 μg/mL aprotinin. Samples were electrophoresed on 10% sodium dodecyl sulfate-polyacrylamide gels and detected using anti-ABCA1 (1:5000 dilution), ABCG1 (1:1000 dilution), ABCG4 (1:1000 dilution), ABCG5 (1:3000 dilution), ABCG8 (1:1000 dilution), or vinculin (1:20,000 dilution) antibodies.
Matsuo M., Ogata Y., Yamanashi Y, & Takada T. (2023). ABCG5 and ABCG8 Are Involved in Vitamin K Transport. Nutrients, 15(4), 998.
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Publication 2023
ABCA1 protein, human ABCG1 protein, human Antibodies Aprotinin Buffers Cells Fluorides leupeptin Phosphates polyacrylamide gels Saline Solution Sodium Chloride Sulfate, Sodium Dodecyl Technique, Dilution Triton X-100 Tromethamine VCL protein, human
2
Antibody Preparation for ABCA1, ABCG4, ABCG5, ABCG1, ABCG8
Mouse anti-ABCA1, rabbit anti-ABCG4, and mouse anti-ABCG5 antibodies were prepared, as described previously [4 (link),8 (link),14 (link)]. Rabbit anti-ABCG1 (sc-20795), anti-ABCG8 (NB400-110), and mouse anti-vinculin (V9131) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), Novus Biologicals (Littleton, CO, USA), and Sigma-Aldrich (St. Louis, MO, USA), respectively. Other chemicals were purchased from Sigma-Aldrich, GE Healthcare (Little Chalfont, UK), Cayman Chemical (Ann Arbor, MI, USA), Wako Pure Chemical Industries (Osaka, Japan), and Nacalai Tesque (Kyoto, Japan).
Matsuo M., Ogata Y., Yamanashi Y, & Takada T. (2023). ABCG5 and ABCG8 Are Involved in Vitamin K Transport. Nutrients, 15(4), 998.
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Publication 2023
ABCA1 protein, human ABCG1 protein, human Anti-Antibodies Antibodies Biological Factors Caimans Mice, House Novus Rabbits Vinculin
3
Quantitative Analysis of Gene Expression
Purification of RNA, preparation of cDNA and RT-qPCR were carried out, as previously described, with transcript levels determined using the comparative Ct method [6 (link),11 (link),42 (link),45 (link)]. The sequences of the primers were: 5′-AGCCATTTTAAAGATAGCTTTCC-3′ and 5′-AAGCTCTGGTTCTTATTCACA-3′ for CD36; 5′-GTCCAATAGGTCCTCCGGGT-3′ and 5′-CCCACCGACCAGTCGAAC-3′ for SRA; 5′-GAGATTTCTCTGTATGGCACC-3′ and 5′-CTGCAAATGAGACACTTTCTC-3′ for LPL; 5′-AGTGGAAACAGTTAATGACCAG-3′ and 5′-GCAGCTGACATGTTTGTCTTC-3′ for ABCA1; 5′-GGTGGACGAAGAAAGGATACAAGACC-3′ and 5′-ATGCCCGTCTCCCTGTATCCA-3′ for ABCG1; 5′-CCTTCAGAACCCACAGAGATCC-3′ and 5′-ACGCTGCATAGCTCGTTCC-3′ for LXR-α; 5-GCTAACAGCGGCTCAAGAACT-3′ and 5′-GGAGCGTTTGTTGCACTGC-3′ for LXR-β; 5′-CAGGAGCCGACTGGCCAATC-3′ and 5′-ACCTTGGCCTGGCATCCTG-3′ for ApoE; and 5-CTTTTGCGTCGCCAGCCGAG-3′ and 5′-GCCCAATACGACCAAATCCGTTGACT-3′ for GAPDH.
O’Morain V.L., Chen J., Plummer S.F., Michael D.R, & Ramji D.P. (2023). Anti-Atherogenic Actions of the Lab4b Consortium of Probiotics In Vitro. International Journal of Molecular Sciences, 24(4), 3639.
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Publication 2023
ABCA1 protein, human ABCG1 protein, human Apolipoprotein E3 apolipoprotein E5 DNA, Complementary GAPDH protein, human Oligonucleotide Primers
4
Western Blot Analysis of Cholesterol Transporters
RAW264.7 cells were cultured on six-well plates with 10% FBS DMEM at a density of 1.0 × 106 cells/well. When the RAW264.7 cells adhered, they were incubated with or without indicated treatment. After the cells were collected, they were washed twice with ice-cold phosphate-buffered saline (PBS). RIPA lysate and protease inhibitor 100:1 were used to extract the protein, and total protein concentration was calculated using a bicinchoninic acid (BCA) protein assay kit (Thermo Scientific, Waltham, MA, USA). The extracted proteins were separated on an 8% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel at 100 V, and the protein was transferred from the gel to polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, MA, USA) at 250 mA for 150 min. The blots were then blocked for 1.5 h at room temperature with skimmed milk powder in 5% bovine serum albumin (BSA). The blots were incubated overnight shaker at 4 °C with mouse anti-ABCA1 (1:500, Abcam, Cambridge, MA, USA), rabbit anti-ABCG1 (1:500, Proteintech, Wuhan, China), rabbit anti-SR-BI (1:1000, Sangon Biotech, Shanghai, China), and rabbit anti-GAPDH (1:1000, Sangon Biotech, Shanghai, China). After rinsing three times for 30 min with TBST, the blots were incubated with horseradish peroxidase (HRP)-labeled goat anti-rabbit IgG (1:1000, Beyotime, Shanghai, China) or HRP-conjugated anti-mouse IgG (1:1000, CST, USA) for 70 min at room temperature. Lastly, the protein expression was detected using a chemiluminescence Western blotting system.
Zhang Z., Qiu Y., Li W., Tang A., Huang H., Yao W., Li H, & Zou T. (2023). Astaxanthin Alleviates Foam Cell Formation and Promotes Cholesterol Efflux in Ox-LDL-Induced RAW264.7 Cells via CircTPP2/miR-3073b-5p/ABCA1 Pathway. Molecules, 28(4), 1701.
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Publication 2023
ABCA1 protein, human ABCG1 protein, human anti-IgG bicinchoninic acid Biological Assay Cells Chemiluminescence Cold Temperature GAPDH protein, human Goat Horseradish Peroxidase IGG-horseradish peroxidase Milk, Cow's Mus Phosphates polyvinylidene fluoride Powder Protease Inhibitors Proteins Rabbits Radioimmunoprecipitation Assay RAW 264.7 Cells Saline Solution SDS-PAGE Serum Albumin, Bovine Tissue, Membrane
5
Quantitative RNA Expression Analysis
Total RNA was extracted using Trizol reagent (Invitrogen, Gaithersburg, MD, USA). The quality and quantity of RNA were determined spectrophotometrically at optical density (OD)260 and OD260/OD280 = 1.8–2.1 using a NanoDrop spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). For qRT-PCR analyses, 1 μg of total RNA was used with a reverse transcription kit (Roche, Shanghai, China) to obtain cDNA according to the manufacturer’s protocol. The specific primers of the mRNAs for mouse ABCA1, ABCG1, SR-BI, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are listed in Table 1. PCR was performed using the following light cycle conditions: 95 °C for 10 min, followed by 40 cycles of 95 °C for 15 s, 56 °C for 30 s. The synthesis of polymerase chain reaction products was monitored by melting curve analysis and agarose gel electrophoresis. The quantitative results for mRNA and circRNA were normalized by glyceraldehyde-3-phosphate dehydrogenase and 18s rRNA, respectively. The relative quantification was calculated using the 2−ΔΔCt method.
Zhang Z., Qiu Y., Li W., Tang A., Huang H., Yao W., Li H, & Zou T. (2023). Astaxanthin Alleviates Foam Cell Formation and Promotes Cholesterol Efflux in Ox-LDL-Induced RAW264.7 Cells via CircTPP2/miR-3073b-5p/ABCA1 Pathway. Molecules, 28(4), 1701.
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Publication 2023
ABCA1 protein, human ABCG1 protein, human Anabolism DNA, Complementary Electrophoresis, Agar Gel Glyceraldehyde-3-Phosphate Dehydrogenases Mus Oligonucleotide Primers Polymerase Chain Reaction Reverse Transcription RNA, Circular RNA, Messenger RNA, Ribosomal, 18S trizol Vision

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Ab52617 by Abcam
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Rneasy mini kit by Qiagen
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The RNeasy Mini Kit is a laboratory equipment designed for the purification of total RNA from a variety of sample types, including animal cells, tissues, and other biological materials. The kit utilizes a silica-based membrane technology to selectively bind and isolate RNA molecules, allowing for efficient extraction and recovery of high-quality RNA.
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Ab18180 by Abcam
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Abca1 by Abcam
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ABCA1 is a membrane protein that functions as a cholesterol efflux pump in the cellular lipid removal pathway. It facilitates the transfer of cellular cholesterol and phospholipids to lipid-poor apolipoproteins, such as apolipoprotein A-I, the major protein component of high-density lipoprotein (HDL) particles.
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Abcg1 by Abcam
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ABCG1 is a membrane-bound protein that belongs to the ATP-binding cassette (ABC) transporter superfamily. It functions as a cholesterol efflux pump, mediating the transport of cholesterol out of cells.
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Anti abcg1 by Abcam
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Anti-ABCG1 is a primary antibody that recognizes the ABCG1 protein. ABCG1 is a member of the ATP-binding cassette (ABC) transporter superfamily and is involved in the regulation of cellular cholesterol homeostasis.

More about "ABCG1 protein, human"

ABCG1 is a member of the ATP-binding cassette (ABC) transporter superfamily, playing a crucial role in cellular cholesterol homeostasis and regulation of cholesterol efflux from cells.
This protein is expressed in various tissues, including macrophages, and its dysregulation has been implicated in the pathogenesis of cardiovascular and metabolic disorders.
Researchers studying ABCG1 can leverage the power of PubCompare.ai, an AI-driven tool that helps identify the most reproducible and effective research protocols from the literature, preprints, and patents.
This streamlines the research process and takes ABCG1 protein studies to new heights by optimizing the selection of experimental methods.
The ABCG1 protein is closely related to the ABCA1 transporter, which is also involved in cellular cholesterol homeostasis.
Antibodies such as Ab52617 and Ab18180 can be used to detect and analyze the expression of ABCG1 in various experimental models.
Techniques like TRIzol reagent and RNeasy Mini Kit can be employed to extract and purify RNA for ABCG1 gene expression analysis, while the High-Capacity cDNA Reverse Transcription Kit can be used for cDNA synthesis.
PVDF membranes and TRIzol can be utilized in Western blot analysis to quantify ABCG1 protein levels.
By leveraging the insights and tools provided by PubCompare.ai, researchers can streamline their ABCG1 protein studies, optimize experimental methods, and advance our understanding of the role of this key transporter in health and disease.