ATP-Binding Cassette, Sub-Family G (WHITE), Member 8 (ABCG8) (N-Term) antibody

Details for Product No. ABIN152894
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Antigen
Synonyms DDBDRAFT_0167690, DDBDRAFT_0191232, DDB_0167690, DDB_0191232, zgc:172358, GBD4, STSL, 1300003C16Rik, AI114946, sterolin-2
Epitope
N-Term
(8)
Reactivity
Human
(51), (37), (31), (25), (25), (25)
Host
Rabbit
(53), (1)
Clonality
Polyclonal
Conjugate
Un-conjugated
(4), (4), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (1)
Application
Western Blotting (WB), Immunocytochemistry (ICC), Immunofluorescence (IF)
(35), (20), (11), (6), (5), (4), (1)
Pubmed 4 references available
Quantity 0.2 mL
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Catalog No. ABIN152894
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Immunogen A synthetic peptide from the N-terminal region of human ABCG8 protein.
Purification affinity purified
Alternative Name ABCG8
Background ATP-binding cassette (ABC) transporter genes are involved in the regulation of theamount of dietary cholesterol retained in the body. ABCG8, expressed at high levels inthe liver and intestine, normally cooperates with ABCG5 to limit intestinal absorption andpromote biliary excretion of sterols. The mutated form of this transporter can lead tosterol accumulation and atherosclerosis or sitosterolemia, a rare autosomal recessivedisorder, characterized by hyperabsorption of sterols and the inability to excrete sterolsinto bile. Alternate Names: anti-ATP binding cassette sub family G (WHITE) member 8 (sterolin 2) antibody,anti-ATP binding cassette sub family G member 8 antibody, anti-MGC142217 antibody,anti-Sterolin 2 antibody, anti-STSL antibody, anti-ATP binding cassette G8 antibody. Related Diseases: Atherosclerosis, Coronary Artery Disease
Gene Symbol: ABCG8
Gene ID 64241, 67470
UniProt Q9H221
Research Area Cancer, Atherosclerosis, Metabolism
Application Notes This ABCG8 antibody is useful for Immunocytochemistry/Immunofluorescence and Western Blot where a doublet is seen at ~67kDa and ~72kDa. Longer exposure times are needed to visualize the wild type ABCG8. For mouse, 100 µg protein (B6/129AKRFVB liver homogenate) loaded with antibody diluted at 1:500. In ICC/IF puncate membrane localization with some weak nuclear reactivity was observed in HEK293 cells.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:250, Western Blot 1:250-1:500
Protocol Western blot protocol specific for ABCG8 Antibody Western Blot Protocol
1. Load ~50 micrograms of lysate (ie: liver membrane) onto an 8 % SDS-PAGE, reducing.
. Block the protein transferred membrane for 30 minutes in blocking buffer [5 % NFDM, 10 mM Tris (pH 7.
6) saline, 0.2 % Tween-20] at room temperature.
. Incubate the membrane in diluted primary anti-ABCG8 [cat# NB 400-117], 1:250 for 1 hour at room temperature.
. Wash the membrane 3x 5 minutes in blocking buffer without milk.
. Incubate the membrane in diluted secondary anti-Rabbit IgG conjugated to HRP.
. Wash the membrane 3x 5 minutes in blocking buffer without milk.
. Wash the membrane for 5 minutes in TBS.
. Develop using West-Pico ECL reagent (Pierce). ICC/IF protocol specific for ABCG8 Antibody Immunocytochemistry ProtocolCulture cells to appropriate density in 35 mm culture dishes or 6-well plates.
. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
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 Avoid freeze-thaw cycles
Storage -20 °C
Storage Comment Aliquot and store at -20 °C or -80 °C.
General Garcia, Wilund, Arca et al.: "Autosomal recessive hypercholesterolemia caused by mutations in a putative LDL receptor adaptor protein." in: Science (New York, N.Y.), Vol. 292, Issue 5520, pp. 1394-8, 2001 (PubMed).

Lu, Lee, Hazard et al.: "Two genes that map to the STSL locus cause sitosterolemia: genomic structure and spectrum of mutations involving sterolin-1 and sterolin-2, encoded by ABCG5 and ABCG8, respectively." in: American journal of human genetics, Vol. 69, Issue 2, pp. 278-90, 2001 (PubMed).

Schmitz, Langmann, Heimerl: "Role of ABCG1 and other ABCG family members in lipid metabolism." in: Journal of lipid research, Vol. 42, Issue 10, pp. 1513-20, 2001 (PubMed).

Mathur, Watt, Field: "Regulation of intestinal NPC1L1 expression by dietary fish oil and docosahexaenoic acid." in: Journal of lipid research, Vol. 48, Issue 2, pp. 395-404, 2007 (PubMed).

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