- PA1-16789 - Invitrogen Antibodies ABCA1 antibody

Submitted references Givinostat-Liposomes: Anti-Tumor Effect on 2D and 3D Glioblastoma Models and Pharmacokinetics.

Taiarol L, Bigogno C, Sesana S, Kravicz M, Viale F, Pozzi E, Monza L, Carozzi VA, Meregalli C, Valtorta S, Moresco RM, Koch M, Barbugian F, Russo L, Dondio G, Steinkühler C, Re F
Cancers 2022 Jun 16;14(12)

CTRP12 ameliorates atherosclerosis by promoting cholesterol efflux and inhibiting inflammatory response via the miR-155-5p/LXRα pathway.

Wang G, Chen JJ, Deng WY, Ren K, Yin SH, Yu XH
Cell death & disease 2021 Mar 10;12(3):254

The Liver X Receptor Is Upregulated in Monocyte-Derived Macrophages and Modulates Inflammatory Cytokines Based on LXRα Polymorphism.

Kim HA, Baek WY, Han MH, Jung JY, Suh CH
Mediators of inflammation 2019;2019:6217548

ATP Binding Cassette Transporter A1 is Involved in Extracellular Secretion of Acetylated APE1/Ref-1.

Lee YR, Joo HK, Lee EO, Cho HS, Choi S, Kim CS, Jeon BH
International journal of molecular sciences 2019 Jun 28;20(13)

PMP22 Regulates Cholesterol Trafficking and ABCA1-Mediated Cholesterol Efflux.

Zhou Y, Miles JR, Tavori H, Lin M, Khoshbouei H, Borchelt DR, Bazick H, Landreth GE, Lee S, Fazio S, Notterpek L
The Journal of neuroscience : the official journal of the Society for Neuroscience 2019 Jul 3;39(27):5404-5418

Calpains Released by T Lymphocytes Cleave TLR2 To Control IL-17 Expression.

Perez J, Dansou B, Hervé R, Levi C, Tamouza H, Vandermeersch S, Demey-Thomas E, Haymann JP, Zafrani L, Klatzmann D, Boissier MC, Letavernier E, Baud L
Journal of immunology (Baltimore, Md. : 1950) 2016 Jan 1;196(1):168-81

Xanthine oxidoreductase is involved in macrophage foam cell formation and atherosclerosis development.

Kushiyama A, Okubo H, Sakoda H, Kikuchi T, Fujishiro M, Sato H, Kushiyama S, Iwashita M, Nishimura F, Fukushima T, Nakatsu Y, Kamata H, Kawazu S, Higashi Y, Kurihara H, Asano T
Arteriosclerosis, thrombosis, and vascular biology 2012 Feb;32(2):291-8

Binding of PDZ-RhoGEF to ATP-binding cassette transporter A1 (ABCA1) induces cholesterol efflux through RhoA activation and prevention of transporter degradation.

Okuhira K, Fitzgerald ML, Tamehiro N, Ohoka N, Suzuki K, Sawada J, Naito M, Nishimaki-Mogami T
The Journal of biological chemistry 2010 May 21;285(21):16369-77

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of ABCA1, using Product # PA1-16789. Samples: 40 µg of total cell post-nuclear lysate of raw macrophages treated with 9-cisretinoic acid and 22R-hydroxycholesterol.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot detection of ABCA1 in mouse peritoneal macrophages using Product # PA1-16789 (Lot L). ECL exposure, 1 min. Lane 4: T09 uninduced lysate Lane 5: T09 induced lysate

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of RAW 264.7 Whole cell Lysate using Product # PA1-16789. Samples: 25 µg whole cell lysate of RAW 264.7 cells treated with T0901317 (lane 1) and untreated (lane 2).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of ABCA1 was performed by loading 20 µg of primary mouse peritoneal macrophage whole cell lysate per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat dry milk in TBST for 1 hour at room temperature. The membrane was probed with an ABCA1 antibody (Product # PA1-16789) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Detection was performed using ECL substrate. Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of ABCA1 in 40 µg total cell lysates of RAW264.9 cells treated with vehicle (-) or 9-cisretinoic acid and 22Rhydroxycholesterol (+). Samples were incubated in ABCA1 polyclonal antibody (Product # PA1-16789.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ABCA1 in mouse peritoneal macrophages. Samples were incubated in ABCA1 polyclonal antibody (Product # PA1-16789). ECL exposure, 1 min. Lane 4: T09 uninduced lysate; Lane 5: T09 induced lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using anti-ABCA1 Polyclonal Antibody (Product # PA1-16789) and a 254 kDa band corresponding to ABCA1 was increased upon THP-1 differentiated to macrophages and LPS treatment in THP-1. Also, the expression was observed to be high in Mouse and Rat Liver when compared to other tissues. Few uncharacterized bands were also observed in liver tissue. Membrane enriched extracts (30 µg lysate) of THP-1 (Lane 1), THP-1 differentiated to macrophages (Lane 2), THP-1 treated with LPS (1 µg/mL for 24hrs) (Lane 3), Mouse Liver (Lane 4), Rat Liver (Lane 5), Mouse Brown Adipose (Lane 6) and Mouse Heart (Lane 7) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gels (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of ABCA1 (green) HEK293T cells. Cells fixed with 4% formaldehyde fixed cells were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with an ABCA1 polyclonal antibody (Product # PA1-16789) at a dilution of 1:100 overnight at 4°C in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with fluorophore-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a Leica DM1000 microscope at 40X magnification. Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of ABCA1 in HepG2 cells were grown to 60% confluency, serum starved for 24 hours, and then treated with 1 µM TO9 for 24 hours prior to being fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. Samples were incubated in ABCA1 polyclonal antibody (Product # PA1-16789) using a dilution of 5 µg/mL overnight at 4 °C followed by anti-rabbit DyLight 488 (Green) with a dilution of 1:500. Alpha tubulin (DM1A) was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of ABCA1 was performed using log phase THP-1 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ABCA1 Polyclonal Antibody (3A1.891.3) (Product # PA1-16789) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of ABCA1 in formalin-fixed paraffin-embedded tissue sections of human prostate cancer were probed for ABCA1 mRNA (ACD RNAScope Probe; Fast Red chromogen, ACD). Samples were incubated in ABCA1 polyclonal antibody (Product # PA1-16789) using a dilution of 1.5 µg/mL with overnight incubation at 4 °C followed by anti-rabbit IgG VisUCyte HRP Polymer Antibody and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of ABCA1 in human prostate epithelium showing luminal and membrane staining. Samples were incubated in ABCA1 polyclonal antibody (Product # PA1-16789).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of ABCA1 in HeLa cells. Samples were incubated in ABCA1 polyclonal antibody (Product # PA1-16789) using a dilution of 1:400. Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunoprecipitation of ABCA1 was performed on THP-1 cells. Antigen-antibody complexes were formed by incubating 400 µg of THP-1 whole cell lysate (in 400 µL volume) with 4 µL of an ABCA1 polyclonal antibody (Product # PA1-16789) overnight at 4°C. The immune complexes were captured on 30 µL of protein G agarose, washed extensively, and eluted with 6X non-reducing Laemmli buffer. Samples were resolved on an 8% SDS-PAGE gel, transferred to a PVDF membrane, and blocked with 5% milk in TBST for 1 hour at room temperature. The membrane was probed with an ABCA1 polyclonal antibody (Product # PA1-16789) at a dilution of 1:500 for overnight at 4øC, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Chemiluminescent detection was performed using ECL substrate. Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 2 Effects of CTRP12 on the expression of cholesterol efflux and uptake markers. A - D THP-1 macrophages were incubated with 50 ug/mL ox-LDL for 48 h, followed by treatment with PBS, LV-NC, or LV-CTRP12 for 72 h. A , B The expression of ABCA1 and ABCG1 was determined by qRT-PCR and western blot. C , D Detection of CD36 and SR-A expression using qRT-PCR and western blot. Data are expressed as the mean +- SD from three independent experiments. **** P < 0.0001; ns not significant.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 3 Involvement of LXRalpha in CTRP12-induced upregulation of ABCA1 and ABCG1 expression. A THP-1 macrophages were treated with 50 ug/mL ox-LDL for 48 h, followed by transfection with PBS, LV-NC, or LV-CTRP12 for 72 h. LXRalpha expression was determined by qRT-PCR and western blot. B After transfection with scrambled siRNA or LXRalpha siRNA for 48 h, cell lysates were immunoblotted with indicated antibodies. C - E THP-1 macrophages were loaded with 50 ug/mL ox-LDL for 48 h, transfected with LXRalpha siRNA for another 48 h and then treated with LV-CTRP12 for 72 h. C The mRNA and protein levels of ABCA1 and ABCG1 were detected by qRT-PCR and western blot, respectively. D , E Representative fluorescent images of NBD-cholesterol burden (x200) and quantitative analyses of cholesterol efflux to apoA-I and HDL. Scale bar = 20 mum. Data are expressed as the mean +- SD from three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 CTRP12-induced upregulation of LXRalpha, ABCA1, and ABCG1 is mediated by miR-155-5p. A THP-1 macrophages were treated with 50 ug/mL ox-LDL for 48 h and then transfected with PBS, LV-NC, or LV-CTRP12 for 72 h, followed by detection of miR-155-5p expression using qRT-PCR. B - D THP-1 macrophages were loaded with 50 ug/mL ox-LDL for 48 h, transfected with miR-155-5p mimic for another 48 h and then transduced with LV-CTRP12 for 72 h. The mRNA and protein levels of LXRalpha, ABCA1, and ABCG1 were detected by qRT-PCR and western blot, respectively. Data are represented as the mean +- SD from three independent experiments. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 8 Effects of CTRP12 on plasma HDL-C levels and RCT in apoE -/- mice. A Plasma levels of TC, TG, HDL-C, and LDL-C were determined using the commercial kits ( n = 10). B Mice were injected intraperitoneally with [ 3 H]-cholesterol-labeled J774 macrophages. The radioactivity in the plasma, liver, and feces were assessed by a liquid scintillation counter ( n = 5). C , D Detection of miR-155-5p, LXRalpha, ABCA1, and ABCG1 expression in the aortas by qRT-PCR and western blot ( n = 10). Data are represented as the mean +- SD. * P < 0.05, **** P < 0.0001; ns not significant.

PA1-16789

Antibody data