ATP-Binding Cassette, Sub-Family A (ABC1), Member 1 (ABCA1) (AA 1100-1300) antibody

Details for Product No. ABIN189744
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Antigen
Synonyms ABC-1, ABCA1, ABC1, CERP, HDLDT1, TGD, Abc1
Epitope
AA 1100-1300
(13), (8), (7), (7), (4), (3), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1)
Reactivity
Hamster, Human, Mouse (Murine), Rat (Rattus)
(64), (63), (37), (25), (12), (3), (2)
Host
Rabbit
(54), (18), (16), (2)
Clonality
Polyclonal
Conjugate
Un-conjugated
(6), (4), (3), (3), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1), (1)
Application
Western Blotting (WB), Chromatin Immunoprecipitation (ChIP), ELISA, Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
(48), (35), (22), (20), (15), (13), (13), (11), (4), (4), (3), (2)
Pubmed 14 references available
Quantity 0.025 mL
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Catalog No. ABIN189744
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Immunogen Partial peptide sequence (within residues 1100-1300) of human ABCA1. Actual immunogen sequence is proprietary information.
Cross-Reactivity (Details) Canine reactivity reported in scientific literature (PMID: 24612239)
Purification affinity purified
Alternative Name ABCA1
Background Mutations in the ABCA1 gene (ATP-binding cassette transporter 1) have been reportedin Tangier disease (TD). TD is an autosomal recessive disorder characterized by anabsence of plasma HDL, cholesterol ester deposition in the reticulo-endothelial systemand deviations in cellular lipid trafficking. ABCA1 mediates the apo-A1 associatedexport of cholesterol and phospholipids from the cell. It is expressed on the plasmamembrane and the Golgi complex, and is regulated by cholesterol flux. Regulation of thecholesterol flux between HDL and macrophages is competitive between ABCA1 andSR-BI. Alternate Names: anti-ABC 1 antibody, anti-ABC Transporter 1 antibody, anti-ABC1 antibody,anti-ATP-binding Cassette 1 antibody, anti-ATP-binding Cassette Transporter 1antibody, anti-CERP antibody, anti-Cholesterol Efflux Regulatory Protein antibody,anti-ABC-1 antibody. Related Diseases: Tangier Disease, Atherosclerosis, Coronary Artery Disease
Gene Symbol: ABCA1
Gene ID 19
UniProt O95477
Application Notes This ABCA1 antibody is useful for Flow Cytometry, Immunocytochemistry/Immunofluorescence (PMID: 21501868), Immunoprecipitation (PMID: 21846716, 21106522), Western blot and Immunohistochemistry on frozen and paraffin embedded sections. In Western blot a band representing ABCA1 is observed at ~220 kDa. Additional non-specific bands are seen at lower molecular weights, but do not interfere with the ABCA1 signal. NOTE: It is important not to boil the sample before loading onto a gel. Boiling can cause aggregation in large proteins, resulting in the proteins inability to enter the gel. Chromatin Immunoprecipitation and ELISA were reported in scientific literature.
Recommended dilutions: Chromatin Immunoprecipitation, ELISA, Flow Cytometry 1:400, Immunocytochemistry/Immunofluorescence 1:100, Immunohistochemistry 1:200, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin 1:200, Immunoprecipitation 1:10-1:500, Western Blot 1:500
Protocol Protocol specific for ABCA1 Antibody Western BlotThe experiment was performed by treating RAW macrophages with 9-cis-retinoic acid and 22R-hydroxycholesterol, known inducers of ABCA1 expression in macrophages. Then total cell post-nuclear lysate (40ug protein) was separated by SDS-PAGE and detected using a 1:1000 dilution of NB 400-105 affinity purified Lot G incubated for 1 hour at room temperature (Lane A). Although there are lower molecular weight bands on the blot, the ABCA1 signal is excellent and gives the expected 3 bands. It is not known why ABCA1 runs as three bands, but it has been found to do so by many researchers. It is probably due to protein modifications such as glycosylation. The antibody was also tested against ABCA1 transiently expressed in 293 cells as an independent test with excellent results.NOTE: An important factor in detecting ABCA1 is in the cell type used. ABCA1 is expressed in very low levels in most cell types. Therefore, ABCA1 expression needs to be induced by using 22-hydroxycholesterol and 9-cis-retinoic acid as ligands for the transcription factor LXR.
1. Without heating at all (leave at room temp for about 15 to 20 minutes with Beta-mercaptoethanol), load 40 µg post-nuclear lysates* to 7.5 % or 4-15 % Tris-HCL SDS gel (Bio-RAD) in sample buffer. Do NOT boil the samples. (NP-40 will not interfere with the running of the protein on SDS-PAGE.)
. Transfer to nitrocellulose membrane at 100V 1hr or 30V overnight.
. Block membrane in 5 % milk in TBS-T for at least 1 hr. Wash with TBS-T 5 minutes.
. Blot with anti-ABCA1 antibody in 3 % milk in TBS-T for 1 hour.
. Wash with TBS-T 3 times, 10 minutes each.
. Blot with donkey anti-rabbit secondary, antibody (Amersham) at 1:2000 in 3 % milk in TBS-T for 1 hour.
. Wash with TBS-T 3 times, 10 minutes each.
. Detect with chemiluminescent reagent (Pierce).
. Expose to X-ray film, 30 sec, 1 min, and 3 min. TBS-T: Tris-buffered-saline with Tween-20See also the specific references mentioned in the datasheet. *Post-nuclear lysate is the result of sonication or dounce homogenization of lysate, centrifugation at low-speed, and the removal of nuclei. The resulting supernatant is called post-nuclear and contains cytosolic and membrane proteins without any of the nuclear components.IHC-FFPE sectionsI. Deparaffinization: A. Treat slides with Xylene: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes. B. Treat slides with 100 % Reagent Alcohol: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.II. Quench Endogenous Peroxidase: A. Place slides in peroxidase quenching solution: 15-30 minutes. To Prepare 200 mL of Quenching Solution: -Add 3 mL of 30 % Hydrogen Peroxide to 200 mL of Methanol. -Use within 4 hours of preparation B. Place slides in distilled water: 2 changes for 2 minutes each. III. Retrieve Epitopes: A. Preheat Citrate
Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer Tris-citrate/Phosphate, pH 7-8, Sodium Azide
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Do not freeze.
Storage 4 °C
Supplier Images
anti-ATP-Binding Cassette, Sub-Family A (ABC1), Member 1 (ABCA1) (AA 1100-1300) antibody Western Blot analysis of ABCA1, using ABIN152883. Samples: 40 ug of total cell post-nuclear lysate of raw macrophages treated with 9-cisretinoic acid and 22R-hydroxycholesterol.
anti-ATP-Binding Cassette, Sub-Family A (ABC1), Member 1 (ABCA1) (AA 1100-1300) antibody (2) Detection of ABCA1 in mouse peritoneal macrophages using ABIN152883 (Lot L). ECL exposure, 1 min. Lane 4: T09 uninduced lysate Lane 5: T09 induced lysate
anti-ATP-Binding Cassette, Sub-Family A (ABC1), Member 1 (ABCA1) (AA 1100-1300) antibody (3) anti-ATP-Binding Cassette, Sub-Family A (ABC1), Member 1 (ABCA1) (AA 1100-1300) antibody (Image 3)
General Burgess, Kiss, Zheng et al.: "Trypsin-sensitive and lipid-containing sites of the macrophage extracellular matrix bind apolipoprotein A-I and participate in ABCA1-dependent cholesterol efflux." in: The Journal of biological chemistry, Vol. 277, Issue 35, pp. 31318-26, 2002 (PubMed).

Murthy, Born, Mathur et al.: "LXR/RXR activation enhances basolateral efflux of cholesterol in CaCo-2 cells." in: Journal of lipid research, Vol. 43, Issue 7, pp. 1054-64, 2002 (PubMed).

Feng, Tabas: "ABCA1-mediated cholesterol efflux is defective in free cholesterol-loaded macrophages. Mechanism involves enhanced ABCA1 degradation in a process requiring full NPC1 activity." in: The Journal of biological chemistry, Vol. 277, Issue 45, pp. 43271-80, 2002 (PubMed).

Fukumoto, Deng, Irizarry et al.: "Induction of the cholesterol transporter ABCA1 in central nervous system cells by liver X receptor agonists increases secreted Abeta levels." in: The Journal of biological chemistry, Vol. 277, Issue 50, pp. 48508-13, 2002 (PubMed).

Wang, Chen, Linsel-Nitschke et al.: "A PEST sequence in ABCA1 regulates degradation by calpain protease and stabilization of ABCA1 by apoA-I." in: The Journal of clinical investigation, Vol. 111, Issue 1, pp. 99-107, 2003 (PubMed).

Escher, Krozowski, Croft et al.: "Expression of sterol 27-hydroxylase (CYP27A1) enhances cholesterol efflux." in: The Journal of biological chemistry, Vol. 278, Issue 13, pp. 11015-9, 2003 (PubMed).

Koldamova, Lefterov, Ikonomovic et al.: "22R-hydroxycholesterol and 9-cis-retinoic acid induce ATP-binding cassette transporter A1 expression and cholesterol efflux in brain cells and decrease amyloid beta secretion." in: The Journal of biological chemistry, Vol. 278, Issue 15, pp. 13244-56, 2003 (PubMed).

Basso, Freeman, Knapper et al.: "Role of the hepatic ABCA1 transporter in modulating intrahepatic cholesterol and plasma HDL cholesterol concentrations." in: Journal of lipid research, Vol. 44, Issue 2, pp. 296-302, 2003 (PubMed).

Terasaka, Hiroshima, Koieyama et al.: "T-0901317, a synthetic liver X receptor ligand, inhibits development of atherosclerosis in LDL receptor-deficient mice." in: FEBS letters, Vol. 536, Issue 1-3, pp. 6-11, 2003 (PubMed).

Sun, Yao, Kim et al.: "Expression of liver X receptor target genes decreases cellular amyloid beta peptide secretion." in: The Journal of biological chemistry, Vol. 278, Issue 30, pp. 27688-94, 2003 (PubMed).

Le Lay, Robichon, Le Liepvre et al.: "Regulation of ABCA1 expression and cholesterol efflux during adipose differentiation of 3T3-L1 cells." in: Journal of lipid research, Vol. 44, Issue 8, pp. 1499-507, 2003 (PubMed).

Khovidhunkit, Moser, Shigenaga et al.: "Endotoxin down-regulates ABCG5 and ABCG8 in mouse liver and ABCA1 and ABCG1 in J774 murine macrophages: differential role of LXR." in: Journal of lipid research, Vol. 44, Issue 9, pp. 1728-36, 2003 (PubMed).

Witting, Maiorano, Davidson: "Ceramide enhances cholesterol efflux to apolipoprotein A-I by increasing the cell surface presence of ATP-binding cassette transporter A1." in: The Journal of biological chemistry, Vol. 278, Issue 41, pp. 40121-7, 2003 (PubMed).

Krimbou, Denis, Haidar et al.: "Molecular interactions between apoE and ABCA1: impact on apoE lipidation." in: Journal of lipid research, Vol. 45, Issue 5, pp. 839-48, 2004 (PubMed).

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