ABCA1 Antibody

Dual RNAscope ISH-IHC: ABCA1 Antibody [NB400-105] - Formalin-fixed paraffin-embedded tissue sections of human prostate cancer were probed for ABCA1 mRNA (ACD RNAScope Probe, catalog # 432291; Fast Red chromogen, ACD catalog # 322360). Adjacent tissue section was processed for immunohistochemistry using rabbit polyclonal (Novus Biologicals catalog # NB400-105) at 1.5ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

Western Blot: ABCA1 Antibody [NB400-105] - Analysis of ABCA1 in total cell lysates of RAW264.9 cells treated with vehicle (-) or 9-cisretinoic acid and 22Rhydroxycholesterol (+). Samples used for this testing were 40 ug of total cell post-nuclear lysate from each group.

Immunocytochemistry/Immunofluorescence: ABCA1 Antibody [NB400-105] - HepG2 cells were grown to 60% confluency, serum starved for 24 hours, and then treated with 1uM 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. The cells were incubated with anti-ABCA1 at 5.0ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 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.

Immunocytochemistry/Immunofluorescence: ABCA1 Antibody [NB400-105] - Untreated HepG2 cells were grown to 60% confluency, and serum starved 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. The cells were incubated with anti-ABCA1 at 5.0ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 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.

Flow Cytometry: ABCA1 Antibody [NB400-105] - ABCA-1 FITC conjugated antibody of human adipose tissue macrophage subsets by flow cytometry. Image from verified customer review.

Western Blot: ABCA1 Antibody [NB400-105] - Prostates of LXR mutant mice accumulate cholesterol esters through inappropriate LXR target genes regulation. Total protein lysates of WT and LXR null mice under normal or high cholesterol diet were analyzed by western blotting with antibodies against ABCA1, LDLR and ACTIN as a loading control (left panel), quantification of ABCA1 and LDLR protein accumulation levels (right panel). * p<0.05, *** p<0.001 in Student's t test. Error bars represent the +/- mean SEM. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pgen.1003483), licensed under a CC-BY licence.

Western Blot: ABCA1 Antibody [NB400-105] - miR-33b regulates ABCA1 and SREBP-1. Analysis for ABCA1, SREBP-1, CPT1a, and AMPK alpha protein levels in the livers of WT, KI+/-, and KI+/+ mice. Representative images are shown. TF2B and beta-actin were used as loading controls. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05312), licensed under a CC-BY licence.

Immunohistochemistry-Paraffin: ABCA1 Antibody [NB400-105] - Association between expression of ABCA1 and survival in ovarian cancer patients. Expression of ABCA1 in 55 ovarian cancer patient samples was determined by IHC in tissue microarray. Representative image of ovarian cancer showing high (left panel) and low (right panel) ABCA1 expression on the cell membrane or cytoplasm (x400). Image collected and cropped by CiteAb from the following publication (//www.clinicalepigeneticsjournal.com/content/7/1/1), licensed under a CC-BY licence.

Flow Cytometry: ABCA1 Antibody [NB400-105] - ABCA1 antibody was tested at 1: 400 in HeLa cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.

Western Blot: ABCA1 Antibody [NB400-105] - Detection of ABCA1 in mouse peritoneal macrophages using NB 400-105 (Lot L). ECL exposure, 1 min. Lane 4: T09 uninduced lysate Lane 5: T09 induced lysate.

Western Blot: ABCA1 Antibody [NB400-105] - miR-33b reduces cellular cholesterol efflux and serum HDL-C levels. Western blotting for ABCA1 (using NB400-105) and ABCG1 proteins in peritoneal macrophages from WT and KI+/+ mice. Representative images are shown. beta-actin was used as the loading control. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05312), licensed under a CC-BY licence.

Immunohistochemistry-Paraffin: ABCA1 Antibody [NB400-105] - Detection of ABCA1 in human prostate epithelium showing luminal and membrane staining.

• Reactivity: Hu, Mu, Rt, Po, Ca, Ch, ChHa, Eq, Ha, Md, Pm, Rb
• Applications: WB, ChIP, ELISA, Flow, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, B/N, Dual ISH-IHC, Func, GS, KD, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

ABCA1 Antibody Summary

• Immunogen: Partial peptide sequence (within residues 1100-1300) of human ABCA1 Antibody [UniProt# O95477]. Actual immunogen sequence is proprietary information.
• Localization: Astrocyte Marker
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: ABCA1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:500
  • Chromatin Immunoprecipitation
  • ELISA
  • Flow Cytometry 1:400
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200
  • Immunoprecipitation 1:10-1:500
  • Block/Neutralize
  • Dual RNAscope ISH-IHC
  • Gel Supershift Assay
  • Knockdown Validated
  • Knockout Validated
  • PCR
• Application Notes: This ABCA1 antibody has been used for Flow Cytometry and ICC (PMID: 21501868), IP (PMID: 21846716; 21106522). Use in Gel supper shift assay reported in scientific literature (PMID 15684432). Use in immunoblotting reported in scientific literature (PMID: 27599291). Use in blocking/ neutralizing reported in scientific literature (PMID: 30821416). Use in PCR reported in scientific literature (PMID: 27406916). Western Blot band representing ABCA1 is observed at approx. 220 kDa. Additional non-specific bands are seen at lower molecular weights, but do not interfere with the ABCA1 signal. Chromatin Immunoprecipitation (PMID 19515742) and ELISA (PMID 18541924) were also reported in scientific literature. Knockdown validation (PMID: 31666189). Knockout validation (PMID: 32273567). In Simple Western only 10-15 uL of the recommended dilution is used per data point. Separated by Charge.
• Theoretical MW: 220 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Raw 264.7 Whole Cell - T0901317 treated Assay Kit

• Control Peptide:

  ABCA1 Protein (NB400-105PEP)

Reactivity Notes

Canine reactivity reported in scientific literature (PMID: 24612239). Rabbit reactivity reported in scientific literature (PMID: 26444796). Chinese Hamster reactivity reported in scientific literature (PMID: 27902765). Primate reactivity reported in scientific literature (PMID: 25440061). Equine reactivity reported in scientific literature (PMID: 26711702).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for ABCA1 Antibody

• ABC1
• ABC-1
• ABC1ATP-binding cassette transporter A1
• ABCA1
• ATP-binding cassette 1
• ATP-binding cassette transporter 1
• ATP-binding cassette, sub-family A (ABC1), member 1
• CERP
• Cholesterol efflux regulatory protein
• EC 2.7.7.8
• EC 2.8.1.8
• EC 3.6.3
• EC 3.6.3.41
• FLJ14958
• HDLDT1
• membrane-bound
• MGC164864
• MGC165011
• TGD
• TGDATP-binding cassette sub-family A member 1

Background

ATP-binding cassette transporter A1 (ABCA1) is a cAMP-dependent and sulfonylurea-sensitive anion transporter belonging to the ATP-binding cassette family. ABCA1 is involved in the regulation of apolipoprotein AI (apoAI)-mediated cholesterol efflux and high-density lipoproteins (HDL) metabolism (1). In the brain, ABCA1 transports cholesterol to apoE, the major CNS apolipoprotein, where it influences motor function and synaptic morphology.

ABCA1 is comprised of 2,261 amino acids with a theoretical molecular weight of 254 kDa and human ABCA1 shares 97% amino acid identity with mouse ABCA1. The general structure of ABCA consists of two transmembrane domains (TMDs) and two nucleotide binding domains (NBDs). ABCA1 is a widely distributed cell-membrane protein with the highest expression found in macrophages. DHHC8 mediated palmitoylation of ABCA1 is essential for its localization to the plasma membrane and expression of mouse ABCA1 (not human) is induced by cAMP analogs (2). ABCA1 is phosphorylated at Ser1042 and Ser2054 by PKA, with Ser2054 being key for regulating phospholipid efflux. Mutations in ABCA1 have been linked to atherosclerosis and the progression of metabolic syndrome phenotypes: high density lipoprotein deficiency type 1 (HDLD1); also known as Tangier disease (TGD), and high density lipoprotein deficiency type 2 (HDLD2); also known as familial hypoalphalipoproteinemia (FHA) (3).

References

1.Oram JF, Lawn RM. (2001) ABCA1. The gatekeeper for eliminating excess tissue cholesterol. J Lipid Res. 42(8):1173-9. PMID: 11483617

2.Singaraja RR, Kang MH, Vaid K, Sanders SS, Vilas GL, Arstikaitis P, Coutinho J, Drisdel RC, El-Husseini Ael D, Green WN, Berthiaume L, Hayden MR. (2009) Palmitoylation of ATP-binding cassette transporter A1 is essential for its trafficking and function. Circ Res. 105(2):138-47. PMID: 19556522

3.Attie AD. (2007) ABCA1: at the nexus of cholesterol, HDL and atherosclerosis. Trends in Biochemical Sciences 32(4):172-9. PMID: 17324574

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ABCA1 Antibody (NB400-105)(356)

Publications using NB400-105

• Brampton C, Pomozi V, Chen LH et al. ABCC6 deficiency promotes dyslipidemia and atherosclerosis Scientific reports Feb 16 2021 [PMID: 33594095] (ICC/IF, Mouse)
• Horie T, Nakao T, Miyasaka Y et al. microRNA-33 maintains adaptive thermogenesis via enhanced sympathetic nerve activity Nature communications Feb 16 2021 [PMID: 33594062] (WB, Mouse)
• Alajbegovic A, Holmberg J, Daoud F, et al. MRTFA overexpression promotes conversion of human coronary artery smooth muscle cells into lipid-laden foam cells Vascular pharmacology Jan 28 2021 [PMID: 33516965] (WB, Human)
• Lv JJ, Wang H, Cui HY, et al. Blockade of Macrophage CD147 Protects Against Foam Cell Formation in Atherosclerosis Frontiers in cell and developmental biology Jan 8 2020 [PMID: 33490072] (WB, ICC/IF, Mouse)
• Palafox-Sanchez V, Ying Z, Royes LFF, Gomez-Pinilla F The interaction between brain and liver regulates lipid metabolism in the TBI pathology Biochimica et biophysica acta. Molecular basis of disease Jan 12 2021 [PMID: 33444711] (WB, Rat)
• Ma C, Zhang J, Yang S et al. Astragalus Flavone Ameliorates Atherosclerosis and Hepatic Steatosis Via Inhibiting Lipid-Disorder and Inflammation in apoE-/- Mice Frontiers in Pharmacology Dec 14 2020 [PMID: 33381046] (WB, Mouse)
• Mukherjee V, Ramakrishna P, Bora S, Kotteazeth S Phytosteroid 28-homobrassinolide targets cholesterol and glucose homeostasis implicating ABCA1 and SREBP role in regulation Steroids Nov 7 2020 [PMID: 33171131] (WB, Human)
  
  Details:
  Western blot analysis performed on HepG2 cells.
• Dodge JC, Jensen EH, Yu J et al. Neutral lipid cacostasis contributes to disease pathogenesis in amyotrophic lateral sclerosis J Neurosci Oct 13 2020 [PMID: 33051352]
• Wang D, Hiebl V, Schachner D et al. Soraphen A enhances macrophage cholesterol efflux via indirect LXR activation and ABCA1 upregulation Biochem. Pharmacol. May 8 2020 [PMID: 32437644] (WB, Human)
• Marshall J, Courage E, Elliott R et al. THP-1 Macrophage Cholesterol Efflux Is Impaired by Palmitoleate Through Akt Activation PLoS One. 2020 May 21 [PMID: 32437392] (WB, Human)
• Beyea, M, Reaume, S, Sawyez, C et al. The Oxysterol 24(S),25-Epoxycholesterol Attenuates Human Smooth Muscle-Derived Foam Cell Formation Via Reduced Low-Density Lipoprotein Uptake and Enhanced Cholesterol Efflux J Am Heart Assoc 2012 (WB, Human)
• Munir MT, Ponce C, Santos JM et al. VD3 and LXR agonist (T0901317) combination demonstrated greater potency in inhibiting cholesterol accumulation and inducing apoptosis via ABCA1-CHOP-BCL-2 cascade in MCF-7 breast cancer cells Mol Biol Rep Oct 1 2020 [PMID: 32990902]
• Zhao D, Li J, Xue C et al. TL1A inhibits atherosclerosis in apoE-deficient mice by regulating the phenotype of vascular smooth muscle cells J. Biol. Chem. Sep 22 2020 [PMID: 32963108] (Mouse)
• Shang Y, Ma C, Zhang J et al. Bifunctional supramolecular nanofiber inhibits atherosclerosis by enhancing plaque stability and anti-inflammation in apoE-/- mice Theranostics Aug 13 2020 [PMID: 32929345] (WB, Mouse)
• Venkat P, Zacharek A, Landschoot-Ward J et al. Exosomes derived from bone marrow mesenchymal stem cells harvested from type two diabetes rats promotes neurorestorative effects after stroke in type two diabetes rats Experimental neurology Sep 2 2020 [PMID: 32889008] (IHC-P, Rat)
• Xia X, Xu Q, Liu M et al. Deubiquitination of CD36 by UCHL1 promotes foam cell formation Cell Death Dis Aug 15 2020 [PMID: 32801299] (IP)
• Gao Q, Wei A, Chen F et al. Enhancing PPAR gamma by HDAC inhibition reduces foam cell formation and atherosclerosis in ApoE deficient mice Pharmacological Research Jul 1 2020 [PMID: 32621955] (WB, Mouse)
• Werkman IL, KOvilein J, de Jonge JC, Baron W Impairing committed cholesterol biosynthesis in white matter astrocytes, but not grey matter astrocytes, enhances in vitro myelination J. Neurochem. Jun 30 2020 [PMID: 32602556] (WB, Rat)
• Pathak P, Helsley RN, Brown AL et al. Small Molecule Inhibition of Gut Microbial Choline Trimethylamine Lyase Activity Alters Host Cholesterol and Bile Acid Metabolism Am. J. Physiol. Heart Circ. Physiol. Apr 24 2020 [PMID: 32330092] (WB, Mouse)
• Hiebl V, Schachner D, Ladurner A et al. Caco-2 Cells for Measuring Intestinal Cholesterol Transport - Possibilities and Limitations Biol Proced Online Apr 11 2020 [PMID: 32308567]
• Salonurmi T, Nabil H, Ronkainen J et Al. 4 beta-Hydroxycholesterol Signals From the Liver to Regulate Peripheral Cholesterol Transporters Front. Pharmacol. Mar 27 2020 [PMID: 32292343] (WB, Rat)
• He Q, Wang F, Honda T et al. Ablation of miR-144 increases vimentin expression and atherosclerotic plaque formation Sci Rep Apr 9 2020 [PMID: 32273567] (KO, Mouse)
• Koga M, Kanaoka Y, Inada K et al. Hesperidin blocks varenicline-aggravated atherosclerotic plaque formation in apolipoprotein E knockout mice by downregulating net uptake of oxidized low-density lipoprotein in macrophages J. Pharmacol. Sci. Feb 28 2020 [PMID: 32173266] (WB, Mouse)
• Bogie Jfj, Grajchen E, Wouters E et Al. Stearoyl-CoA desaturase-1 impairs the reparative properties of macrophages and microglia in the brain J. Exp. Med. May 4 2020 [PMID: 32097464] (Flow, Mouse)
• Matsuda J, Takahashi A, Takabatake Y et al. Metabolic effects of RUBCN/Rubicon deficiency in kidney proximal tubular epithelial cells Autophagy Jan 16 2020 [PMID: 31944172]
• Chen Y, Guo KM, Nagy T, Guo TL Metabolomics study of the prefrontal cortex in a rat model of attention deficit hyperactivity disorder reveals the association between cholesterol metabolism disorder and hyperactive behavior Biochem. Biophys. Res. Commun. Dec 18 2019 [PMID: 31864712] (WB, Rat)
• Koga M, Kanaoka Y, Okamoto M et al. Varenicline aggravates atherosclerotic plaque formation in nicotine-pretreated ApoE knockout mice due to enhanced oxLDL uptake by macrophages through downregulation of ABCA1 and ABCG1 expression J. Pharmacol. Sci. Nov 14 2019 [PMID: 31771811] (WB, Mouse)
• Du Y, Li X, Su C et al. Butyrate protects against high-fat diet-induced atherosclerosis via up-regulating ABCA1 expression in apolipoprotein E-deficiency mice Br. J. Pharmacol. Nov 25 2019 [PMID: 31769014]
• Pagano S, Magenta A, D'Agostino M et al. Anti-ApoA-1 IgGs in Familial Hypercholesterolemia Display Paradoxical Associations with Lipid Profile and Promote Foam Cell Formation J Clin Med Nov 21 2019 [PMID: 31766415] (WB, Human)
• Kaymak I, Maier CR, Schmitz W et al. Mevalonate pathway provides ubiquinone to maintain pyrimidine synthesis and survival in p53-deficient cancer cells exposed to metabolic stress Cancer Res. Nov 19 2019 [PMID: 31744820] (WB, Human, Mouse)
• Xiao L, Luo G, Guo X et al. Macrophage iron retention aggravates atherosclerosis: Evidence for the role of autocrine formation of hepcidin in plaque macrophages Biochim Biophys Acta Mol Cell Biol Lipids 2019 Oct 27 [PMID: 31666189] (ICC/IF, WB, KD, Human, Mouse)
• Smith TKT, Kahiel Z, LeBlond ND et al. Characterization of Redox-Responsive LXR-Activating Nanoparticle Formulations in Primary Mouse Macrophages Molecules Oct 18 2019 [PMID: 31635211] (WB, Mouse)
• Xu Y, Xu Y, Zhu Y et al. Macrophage miR-34a Is a Key Regulator of Cholesterol Efflux and Atherosclerosis Mol. Ther. 2019 Sep 12 [PMID: 31604677]
• Hennessy EJ, van Solingen C, Scacalossi KR et al. The long noncoding RNA CHROME regulates cholesterol homeostasis in primate Nat Metab Jan 1 2019 [PMID: 31410392] (WB, Human)
• Zhou Y, Bazick H, Miles JR et al. A neutral lipid-enriched diet improves myelination and alleviates peripheral nerve pathology in neuropathic mice Exp. Neurol. Aug 3 2019 [PMID: 31386828] (WB, IHC-Fr, Mouse)
• Paul A, Lydic TA, Hogan R, Goo YH Cholesterol Acceptors Regulate the Lipidome of Macrophage Foam Cells Int J Mol Sci Aug 2 2019 [PMID: 31382484] (WB, Mouse)
• Mukhamedova N, Hoang A, Dragoljevic D et al. Exosomes containing HIV protein Nef reorganize lipid rafts potentiating inflammatory response in bystander cells PLoS Pathog. Jul 1 2019 [PMID: 31344124] (ICC/IF, Human, Mouse)
• Koyama S, Horie T, Nishino T et al. Identification of Differential Roles of MicroRNA-33a and -33b During Atherosclerosis Progression With Genetically Modified Mice J Am Heart Assoc Jul 2 2019 [PMID: 31242815] (IHC-Fr, Mouse)
• Rege SD, Luiz R, Tsai B et al. Brain Trauma Disrupts Hepatic Lipid Metabolism: Blame it on Fructose? Mol Nutr Food Res May 13 2019 [PMID: 31087499] (WB, Rat)
• Melendez-Avalos A, Sainz-Espunes T, Castrillon-Rivera LE et al. PMP22 regulates cholesterol trafficking and ABCA1-mediated cholesterol efflux J. Neurosci. May 6 2019 [PMID: 31061090] (WB, Mouse)
• Tiwari P, Blank A, Cui C, Schoenfelt KQ et al Metabolically activated adipose tissue macrophages link obesity to triple-negative breast cancer. J Exp Med. 2019 Jun 3 [PMID: 31053611] (Flow, Human)
  
  Details:
  Citation using the DyLight 650 version of this antibody.
• Liao S, Zhou Q, Zhang Y. Elastic aortic wrap reduced aortic stiffness by partially alleviating the impairment of cholesterol efflux capacity in pigs J Diabetes Metab Disord Aug 7 2018 [PMID: 30918842] (WB, Porcine)
• Storti F, Klee K, Todorova V et al. Impaired ABCA1/ABCG1-mediated lipid efflux in the mouse retinal pigment epithelium (RPE) leads to retinal degeneration Elife Mar 13 2019 [PMID: 30864945] (WB, ICC/IF, Mouse)
• Margaryan NV, Hazard-Jenkins H, Salkeni MA et al. The Stem Cell Phenotype of Aggressive Breast Cancer Cells Cancers (Basel) Mar 8 2019 [PMID: 30857267] (IHC, Human)
• Cabou C, Honorato P, Briceno L et al. Pharmacological inhibition of the F1 -ATPase/P2Y1 pathway suppresses the effect of apolipoprotein A1 on endothelial nitric oxide synthesis and vasorelaxation Acta Physiol (Oxf) Mar 1 2019 [PMID: 30821416] (B/N, Mouse)
• Nakazeki F, Tsuge I, Horie T et al. MiR-33a is a therapeutic target in SPG4-related hereditary spastic paraplegia human neurons Clin. Sci. Feb 28 2019 [PMID: 30777884] (WB, Human)
• Tang S, Tabet F, Cochran BJ et al. Apolipoprotein A-I enhances insulin-dependent and insulin-independent glucose uptake by skeletal muscle Sci Rep Feb 4 2019 [PMID: 30718702] (WB, Human)
• Vaidya M, Jentsch JA, Peters S et al. Regulation of ABCA1-mediated cholesterol efflux by sphingosine-1-phosphate signalling in macrophages. J. Lipid Res. Jan 17 2019 [PMID: 30655318] (WB, Mouse)
• Ramon-Vazquez A, de la Rosa JV, Tabraue C et al. Common and Differential Transcriptional Actions of Nuclear Receptors LXRa and LXRb in macrophages. Mol. Cell. Biol. Jan 2 2019 [PMID: 30602495] (WB, Mouse)
• Moon SH, Huang CH, Houlihan SL et al. p53 Represses the Mevalonate Pathway to Mediate Tumor Suppression Cell Dec 14 2018 [PMID: 30580964] (WB, Human)
• Torres-Adorno AM, Vitrac H, Qi Y et al. Eicosapentaenoic acid in combination with EPHA2 inhibition shows efficacy in preclinical models of triple-negative breast cancer by disrupting cellular cholesterol efflux. Oncogene. Nov 20 2018 [PMID: 30459358] (Human)
• Zinkhan EK, Yu B, McKnight R. Uteroplacental Insufficiency Impairs Cholesterol Elimination in Adult Female Growth-Restricted Rat Offspring Fed a High-Fat Diet. Reprod Sci. Nov 19 2018 [PMID: 30453824] (Rat)
• Han Q, Yeung SC, Ip MSM, Mak JCW. Dysregulation of cardiac lipid parameters in high-fat high-cholesterol diet-induced rat model. Biochem. Pharmacol. Nov 9 2018 [PMID: 30428911] (WB, Rat)
• O'Reilly ME, Kajani S, Ralston JC et al. Nutritionally Derived Metabolic Cues Typical of the Obese Microenvironment Increase Cholesterol Efflux Capacity of Adipose Tissue Macrophages. Mol Nutr Food Res. Nov 9 2018 [PMID: 30411491] (WB, Mouse)
• Jin P, Bian Y, Wang K et al. Homocysteine accelerates atherosclerosis via inhibiting LXRa-mediated ABCA1/ABCG1-dependent cholesterol efflux from macrophages Life Sciences Dec 1 2018 [PMID: 30393020] (WB, Human)
• Yuan C, Guo X, Zhou Q et al. OAB-14, a bexarotene derivative, improves Alzheimer's disease-related pathologies and cognitive impairments by increasing beta-amyloid clearance in APP/PS1 mice. Biochim Biophys Acta Mol Basis Dis 2019 Jan [PMID: 30389579] (Mouse)
• Zhong S, Li L, Zhang YL et al. Acetaldehyde dehydrogenase 2 interactions with LDLR and AMPK regulate foam cell formation. J. Clin. Invest. Oct 30 2018 [PMID: 30375985] (WB, Mouse)
• Nishino T, Horie T, Baba O et al. SREBF1/MicroRNA-33b Axis Exhibits Potent Effect on Unstable Atherosclerotic Plaque Formation In Vivo Arteriosclerosis, Thrombosis, and Vascular Biology [PMID: 30354203] (WB, Mouse)
• Liao S, Wu H, Chen R. Apolipoprotein A1 mimetic peptide ATI-5261 reverses arterial stiffness at late pregnancy and early postpartum in a COMT-/- mouse model of preeclampsia. Clin Hypertens Sep 15 2018 [PMID: 30237900] (WB, Mouse)
• de Neergaard R, Nielsen JE, Jorgensen A et al. Positive association between cholesterol in human seminal plasma and sperm counts: results from a cross-sectional cohort study and immunohistochemical investigations. Andrology Sep 4 2018 [PMID: 30182437] (IHC, Human)
• Wang X, Luo J, Li N et al. E3317 promotes cholesterol efflux in macrophage cells via enhancing ABCA1 expression. Biochem. Biophys. Res. Commun. Sep 26 2018 [PMID: 30173890] (WB, Human)
• Qian K, Feng L, Sun Y et al. Overexpression of Salusin-a Inhibits Vascular Intimal Hyperplasia in an Atherosclerotic Rabbit Model. BioMed Research International Jul 12 2018 [PMID: 30105261] (WB, Rabbit)
• Lyssenko NN, Haider N, Picataggi A et al. Directional ABCA1-mediated cholesterol efflux and apoB-lipoprotein secretion in the retinal pigment epithelium J. Lipid Res. Aug 3 2018 [PMID: 30076206] (IHC, WB, Human)
• Wang L, Eftekhari P, Schachner D et al. Novel interactomics approach identifies ABCA1 as direct target of evodiamine, which increases macrophage cholesterol efflux. Sci Rep. Jul 23 2018 [PMID: 30038271] (WB, Human)
• Chernick D, Ortiz-Valle S, Jeong A et al. HDL Mimetic Peptide 4F Mitigates AB-Induced Inhibition of ApoE Secretion and Lipidation in Primary Astrocytes and Microglia. J. Neurochem. Jul 20 2018 [PMID: 30028014] (WB, Mouse)
• Sonett J, Goldklang M, Sklepkiewicz P et al. A critical role for ABC transporters in persistent lung inflammation in the development of emphysema after smoke exposure. FASEB J 2018 Jun 15 [PMID: 29906247] (Mouse)
• Zinkhan EK, Yu B, Schlegel A. Prenatal Exposure to a Maternal High Fat Diet Increases Hepatic Cholesterol Accumulation in Intrauterine Growth Restricted Rats in Part Through MicroRNA-122 Inhibition of Cyp7a1 Front Physiol May 29 2018 [PMID: 29896121] (WB, Rat)
• Wang D, Hiebl V, Ladurner A et al. 6-Dihydroparadol, a Ginger Constituent, Enhances Cholesterol Efflux from THP-1-derived Macrophages Mol Nutr Food Res May 26 2018 [PMID: 29802792] (WB, Human)
• Feng T, Liu P, Wang X et al. SIRT1 activator E1231 protects from experimental atherosclerosis and lowers plasma cholesterol and triglycerides by enhancing ABCA1 expression Atherosclerosis Apr 30 2018 [PMID: 29787963] (WB, Mouse)
• Ishigami M, OgaSawara F, Nagao K et al. Temporary sequestration of cholesterol and phosphatidylcholine within extracellular domains of ABCA1 during nascent HDL generation. Sci Rep Apr 18 2018 [PMID: 29670126] (WB, Human)
• Xie Z, Wang X, Liu X et al. Adipose-Derived Exosomes Exert Proatherogenic Effects by Regulating Macrophage Foam Cell Formation and Polarization. J Am Heart Assoc. Mar 3 2018 [PMID: 29502100] (WB, Mouse)
• Ahn SB, Jun DW, Jang K et al. Duodenal Niemann-Pick C1-like 1 expression was negatively correlated with liver X receptor expression in nonalcoholic fatty liver disease. Korean J Intern Med 2018 Feb 23 [PMID: 29466845]
• Casali BT, Reed-Geaghan EG, Landreth GE. Nuclear receptor agonist-driven modification of inflammation and amyloid pathology enhances and sustains cognitive improvements in a mouse model of Alzheimer's disease. J Neuroinflammation. 2018 Feb 15 [PMID: 29448961] (WB, Mouse)
• Sallam T, Jones M, Thomas BJ, Wu X. Transcriptional regulation of macrophage cholesterol efflux and atherogenesis by a long noncoding RNA. Nat. Med. 2018 Feb 12 [PMID: 29431742] (WB)
• Li Zhiqiang, Kabir Inamul, Tietelman Gladys et al. Sphingolipid de novo biosynthesis is essential for intestine cell survival and barrier function. Cell Death & Disease 2018 Feb 7 [PMID: 29415989] (WB, Mouse)
• Li Y, Shen S, Ding S, Wang L. Toll-like receptor 2 downregulates the cholesterol efflux by activating the nuclear factor‑kappa B pathway in macrophages and may be a potential therapeutic target for the prevention of atherosclerosis Experimental and Therapeutic Medicine 2017 Oct 31 [PMID: 29375683] (WB, Mouse)
• Milligan S, Martin GG, Landrock D et al. Ablating both Fabp1 and Scp2/Scpx (TKO) induces hepatic phospholipid and cholesterol accumulation in high fat-fed mice. Biochim Biophys Acta Mol Cell Biol Lipids 2018 Mar [PMID: 29307784] (Mouse)
• Widenmaier SB, Snyder NA, Nguyen TB et al. NRF1 Is an ER Membrane Sensor that Is Central to Cholesterol Homeostasis Cell. 2017 Nov 16 [PMID: 29149604] (WB)
• Doran AC, Ozcan L, Cai B et al. CAMKIIg suppresses an efferocytosis pathway in macrophages and promotes atherosclerotic plaque necrosis. J. Clin. Invest. 2017 Nov 01 [PMID: 28972541] (WB)
• Storti F, Raphael G, Griesser V et al. Regulated efflux of photoreceptor outer segment-derived cholesterol by human RPE cells Exp. Eye Res. 2017 Sep 21 [PMID: 28943268] (ICC/IF, Human)
• Pelletier RM, Akpovi CD, Chen L et al. Cholesterol metabolism and Cx43, Cx46, and Cx50 gap junction protein expression and localization in normal and diabetic and obese ob/ob and db/db mouse testes Am J Physiol Endocrinol Metab. 2017 Dec 31 [PMID: 28851737] (WB, Mouse)
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  Details:
  This publication used the DL488 conjugated form of this antibody (NB400-105G)
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• Koldamova, R et al. The Liver Receptor Lig T0901317 Decreases Amyloid Beta Production in Vitro in a Mouse Model of Alzheimer';s Disease. The Journal of Biological Chemistry 280(6): 4079-4088. 2005 [PMID: 15557325] (WB, Mouse)
• Dove DE, Su YR, Zhang W, Jerome WG, Swift LL, Linton MF, Fazio S. ACAT1 deficiency disrupts cholesterol efflux and alters cellular morphology in macrophages. Arterioscler Thromb Vasc Biol;25(1):128-34. 2005 Jan. [PMID: 15499044]
• O'Connell BJ, Denis M, Genest J. Cellular physiology of cholesterol efflux in vascular endothelial cells. Circulation;110(18):2881-8. 2004 Nov 2. [PMID: 15492319]
• Bared SM, Buechler C, Boettcher A, Dayoub R, Sigruener A, Grandl M, Rudolph C, Dada A, Schmitz G. Association of ABCA1 with syntaxin 13 and flotillin-1 and enhanced phagocytosis in tangier cells. Mol Biol Cell;15(12):5399-407. 2004 Dec. [PMID: 15469992]
• Zeng, L et al. Sterol-responsive Element-binding Protein (SREBP) 2 Down-regulates ATP-binding Cassette Transporter A1 in Vascular Endothelial Cells: A NOVEL ROLE OF SREBP IN REGULATING CHOLESTEROL METABOLISM. J Biol Chem 279: 48801-48807. 2004 [PMID: 15358760] (WB, Human)
• Denis, M et al. Characterization of Oligomeric Human ATP Binding Cassette Transporter A1. The Journal of Biological Chemistry 279(40):41529-41536. 2004 [PMID: 15280376] (IP, Human)
• Ando H, Tsuruoka S, Yamamoto H, Takamura T, Kaneko S, Fujimura A. Effects of pravastatin on the expression of ATP-binding cassette transporter A1. Exp Ther. 311(1):420-5. 2004 Oct. [PMID: 15210833]
• Brown III, J et al. Differential Expression of Cholesterol Hydroxylase in Alzheimer's Disease. The Journal of Biological Chemistry 279(33): 34674-34681. 2004 [PMID: 15148325] (WB, Human)
• Agassandian M, Mathur SN, Zhou J, Field FJ, Mallampalli RK. Oxysterols trigger ABCA1-mediated basolateral surfactant efflux. Am J Respir Cell Mol Biol;31(2):227-33. 2004 Aug. [PMID: 15039140]
• Sahoo, D et al. ABCA1-dependent lipid effulux to apolipoprotein A-1 mediates HDL particle formation decreases VLDL secretion from murine hepatocytes. Journal of Lipid Research 45: 1122-1131. 2004 [PMID: 14993246] (WB, Rat)
• Field, F et al. LXR/RXR lig activation enhances basolateral efflux of beta-sitosterol in CaCo-2 cells. Journal of Lipid Research 45:905-913. 2004 [PMID: 14993242] (WB, Human)
• Krimbou L, Denis M, Haidar B et al. Molecular interactions between apoE ABCA1: impact on apoE lipidation. Journal of Lipid Research 45:839-848. 2004 [PMID: 14754908] (IP, Human)
• Gaus K, Kritharides L, Schmitz G, Boettcher A, Drobnik W, Langmann T, Quinn CM, Death A, Dean RT, Jessup W. Apolipoprotein A-1 interaction with plasma membrane lipid rafts controls cholesterol export from macrophages. FASEB J;18(3):574-6. 2004 Mar. [PMID: 14734645]
• Haidar, B et al. Apolipoprotein A-I Activates Cellular cAMP Signaling through the ABCA1 Transporter. J Biol Chem 279: 9963-9969. 2004 [PMID: 14701824] (IP, Human)
• Gaus, K et al. Visualizing lipid structure raft domains in living cells with two- photon microscopy. PNAS 100(26): 15554-15559. 2003 [PMID: 14673117] (ICC/IF, Mouse, Human)
• Denis, M et al. Molecular Cellular Physiology of Apolipoprotein A-I Lipidation by the ATP-binding Cassette Transporter A1 (ABCA1). Journal of Biological Chemistry 279(9):7384-7394. 2004 [PMID: 14660648] (IP, Human)
• Su, Y et al. Macrophage Apolipoprotein A-I Expression Protects against Atherosclerosis in ApoE-Deficient Mice Up-regulates ABC Transporters. Molecular Therapy 0:1-8. 2003 [PMID: 14529830] (WB, Mouse)
• Zager RA, Johnson AC, Hanson SY. Proximal tubular cholesterol loading after mitochondrial, but not glycolytic, blockade. Am J Physiol Renal Physiol;285(6):F1092-9. 2003 Dec. [PMID: 12952856]
• Bortnick, A. Identification characterization of rodent ABCA1 in isolated type II pneumocytes. Am J Physiol Lung Cell Mol Physiol 285: L869-L878. 2003 [PMID: 12909583] (WB, Human)
• Wagner, B. Promoter-Specific Roles for Liver X Receptor/Corepressor Complexes in the Regulation of ABCA1 SREBP1 Gene Expression. Molecular Cellular Biology, pp 5780-5789. 2003Aug. [PMID: 12897148] (WB, Mouse)
• Witting, S et al. Ceramide Enhances Cholesterol Efflux to Apolipoprotein A-I by Increasing the Cell Surface Presence of ATP-binding Cassette Transporter A1. Journal of Biological Chemistry 278(41):40121-40127. 2003 [PMID: 12890677] (WB, Mouse)
• Khovidhunkit, W et al. Endotoxin down-regulates ABCG5 ABCG8 in mouse liver ABCA1 ABCG1 in J774 murine macrophages: defferential role of LXR. Journal of Lipid Research 44: 1728-1736. 2003 [PMID: 12777468] (WB, Mouse)
• Le Lay, S et al. Regulation of ABCA1 expression cholesterol efflux during adipose differentiation of 3T3-L1 cells, J. Lipid Res 44(8): p 1499-1507. 2003 [PMID: 12754274] (WB, Mouse)
• Sun, Y. Expression of Liver X Receptor Target Genes Decreases Cellular Amyloid beta Peptide Secretion. The Journal of Biological Chemistry. Vol.278, No.30 July 25,pp. 27688-27694. 2003 [PMID: 12754201] (WB, Mouse)
• Terasaka, N et al. T-0901317, a synthetic liver X receptor lig, inhibits development of atherosclerosis in LDL receptor-deficient mice. FEBS Letters 536: 6-11. 2003 [PMID: 12586329] (IHC-P, Mouse)
• Basso, F et al. Role of the hepatic ABCA1 transporter in modulating intrahepatic cholestrol plasma HDL cholestrol concentrations. Journal of Lipid Research 44:296-302. 2003 [PMID: 12576511] (WB, Mouse)
• Koldamova, R et al. 22R-Hydroxycholestrol 9-cis-Retinoic Acid Induce ATP-binding Cassette Transporter A1 Expression Cholesterol Efflux in Brain Cells Decrease Amyloid beta Secretion. Journal of Biological Chemistry 278(15):13244-13256. 2003 [PMID: 12547833] (WB, IHC, ICC/IF, Rat)
• Escher, G et al. Expression of Sterol 27-Hydroxylase (CYP27A1) Enhances Cholesterol Efflux. Journal of Biological Chemistry 278(13):11015-11019. 2003 [PMID: 12531903] (WB, Human, Hamster)
• Wang, N et al. A PEST sequence in ABCA1 regulates degradation by calpain protease stabilization of ABCA1 by apoA-I. Journal of Clinical Investigation111(1): 99-107. 2003 [PMID: 12511593] (WB, IP, Human)
• Johnson AC, Yabu JM, Hanson S et al. Experimental glomerulopathy alters renal cortical cholesterol, SR-B1, ABCA1, and HMG CoA reductase expression Am J Pathol. 2003 Jan [PMID: 12507911] (WB, Mouse)
• Haidar, B et al. camp induces ABC1 phosphorylation activity promotes cholesterol efflux from fibroblasts. Journal of Lipid Research 43(12):2087-2094. 2002 [PMID: 12454270] (IP, Human)
• Reddy ST, Hama S, Ng C, Grijalva V, Navab M, Fogelman AM. ATP-binding cassette transporter 1 participates in LDL oxidation by artery wall cells. Arterioscler Thromb Vasc Biol;22(11):1877-83. 2002 Nov 1. [PMID: 12426219]
• Fukumoto, H et al. Induction of the Cholesterol Transporter ABCA1 in Central Nervous System Cells by Liver X Receptor Agonists Increases Secreted A Beta Levels. Journal of Biological Chemistry 277(50):48508-48513. 2002 [PMID: 12384498] (WB, Mouse, Human)
• Feng, B et al. ABCA1-mediated Cholestrol Efflux Is Defective in Free Cholesterol-loaded Macrophages. Journal of Biological Chemistry 277(45):43271-73280. 2002 [PMID: 12215451] (WB, Mouse)
• Murthy, S et al. LXR/RXR activation enhances basolateral efflux of cholesterol in CaCo-2 cells. J Lipid Research 43: 1054-1064. 2002 [PMID: 12091489] (WB, Human)
• Burgess, JW et al. Trypsin-sensitive lipid-containing sites of the macrophage extracellular matrix bind apolipoprotein A-I participate in the ABCA-1-dependent cholesterol efflux. JBC Papers in Press 277(35): 31318-31326. 2002 [PMID: 12050168] (WB, Mouse, Human)

Reviews for ABCA1 Antibody (NB400-105) (6)

Average Rating: 4.3

(Based on 6 reviews)

We have 6 reviews tested in 4 species: Human, Mouse, Rat, Other.

reviewed by: Jahangir Iqbal WB Mouse 11/16/2015

Summary

• Application: Western Blot
• Sample Tested: mouse liver
• Species: Mouse
• Lot: AF-2

Blocking

• Blocking Details: Membranes were blocked in 5% BSA in TBST for 1 h at room temperature

Primary Anitbody

• Dilution Ratio: Membranes were immunoblotted with anti-ABCA1 antibody 1:2000 dilution in 2.5% BSA in TBST overnight at 4 oC

Secondary Antibody

• Secondary Description: Membranes were incubated with secondary antibody conjugated to stabilized peroxidase for 1 hour at room temperature
• Secondary Manufacturer Cat#: Thermo Scientific # 32460
• Secondary Concentration: dilution 1:15,000

Details

• Detection Notes: Bands were visualized used chemiluminescence according to the manufacturer’s instructions

Comments

• Comments: Samples were not boiled and 2X BME was added.

reviewed by: Anonymous WB Mouse 12/23/2014

Summary

• Application: Western Blot
• Sample Tested: See PMID 23519696
• Species: Mouse

Blocking

• Blocking Details: See PMID 23519696

Details

• Detection Notes: See PMID 23519696

Comments

• Comments: Independent review from antibodyresource.comPublished in PMID 23519696

reviewed by: Angeliki Chroni IF Human 11/03/2014

Summary

• Application: Immunofluorescence
• Sample Tested: SK-N-SH human neuroblastoma cells
• Species: Human

reviewed by: Anonymous WB Rat 10/15/2012

Summary

• Application: Western Blot
• Sample Tested: Cardiomyocytes from newborn rats
• Species: Rat

Blocking

• Blocking Details: PBS Milk 5%, 1 hour, room temperature

Primary Anitbody

• Dilution Ratio: 1000 = dilution, 1 hour, room temperature, PBS without tween and 2% Milk

Secondary Antibody

• Secondary Description: goat anti rabbit IgG HRP
• Secondary Manufacturer Cat#: Amersham NA934V
• Secondary Concentration: 3000 diluted

Details

• Detection Notes: ECL super plus Amersham RPN2132

reviewed by: Cynthia Hong WB Mouse 09/27/2011

Summary

• Application: Western Blot
• Sample Tested: Mouse
• Species: Mouse

reviewed by: Anonymous WB Other 06/15/2009

Summary

• Application: Western Blot
• Sample Tested: Rat astrocyte, Human Astrocyte, and Raw Mouse Neuron, Sample Amount: 25ug to 50ug
• Species: Other

Blocking

• Blocking Details: Blocking Buffer: 5% Milk w/ TBST, Blocking Time: 1 hour, Blocking Temp: room temperature

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1/1000, Primary Ab Incubation Time: 3 hours, Primary Ab Incubation Temp: room temperature

Secondary Antibody

• Secondary Description: Secondary Ab: anti-Rabbit IgG HRP, Secondary Ab Dilution Ratio: 1/1000

Details

• Detection Notes: Exposure Time: 1 hour, Positive Control: HC/RA, Specific Bands: 200 kDa

FAQs for ABCA1 Antibody (NB400-105). (Showing 1 - 10 of 12 FAQs).

1. For Western blot protocol for ABCA1 antibody NB400-105, why does it suggest treating with beta-ME while not heating?
   • For some membrane proteins it is not recommended to boil the samples. You may incubate them at room temperature or at 37C in SDS sample buffer with BME. However, from personal experience, if you boil the samples you will not see the protein on the blot. I do not think the exact reason is known.
2. Do you have a protocol for Flow cytometry analysis with this ABCA1 antibody (NB400-105)?
   • Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.
3. We purchased this antibody a few weeks ago, and we by mistake stored it at -20 degrees Celsius. Is there possible the antibody still works?
   • Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.
4. I am interested in looking at this marker on monocytes by flow cytometry. I noticed that a directly conjugated 488 ab is available, and that your flow cytometry protocol calls for methanol fixation of the cells. Is the fixation required for antibody binding, or may it be used on cells that have not been methanol fixed? I would like to retain other markers.
   • Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.
5. To do Western blot for the determination of ABCA1 protein level, do I need to order NB400-105 or NB400-105B? Which one do you recommend if we also order the secondary antibody catalog # NB730-H?
   • The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.
6. We purchased NB400-105, Lot# Z5, but the datasheet (approx. 450kDa) was differ from your website (approx. 220kDa) for size of WB. Which molecular weight is correct?
   • ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.
7. Does this product contain any BSA?
   • No, this antibody does not have any BSA in its formulation.
8. I bought ABCA1 antibody (NB400-105) from your company recently and used it in Flow cytometry. I want to know the binding site of ABCA1 and the antibody; is it within the cell or outside? Also, do I need to change the cytomembrane's permeabilization before my test?
   • The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.
9. Is post-nuclear lysate is necessary? Can't I lysate my cells with RIPAbuffer (#R0278 sigma)?
   • The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.
10. Why shouldn't I boil the samples? is it OK to boil them if using RIPAbuffer?
    • To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.
11. Can i use DTT instead of Beta- mercaptoethanol?
    • Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.
12. Have you tested abca1 antibody(catalog # NB400-105) on hepatic Abca1 expression?
    • We do not currently have any testing data where we have used NB400-105 with hepatic samples; all of our testing data for NB400-105 can be seen on the product page for this antibody. Should your customer be interested, we do show murine liver data with the following product: NB100-2068.
Show All 12 FAQs.

Bioinformatics

• Gene Symbol: ABCA1
• Entrez:
  • 19(Human)
  • 11303(Mouse)
  • 313210(Rat)
• Uniprot:
  • O95477(Human)