Solute Carrier Family 7, (Cationic Amino Acid Transporter, Y+ System) Member 11 (SLC7A11) (N-Term), (AA 1-50) antibody

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Antigen
Synonyms SLC7A11, DKFZp468E122, CCBR1, xCT, 9930009M05Rik, AI451155, sut
Epitope
N-Term, AA 1-50
(20), (9), (6), (1), (1)
Reactivity
Human, Mouse (Murine), Rat (Rattus)
(39), (20), (15), (13), (12), (11), (1)
Host
Rabbit
(39), (6)
Clonality
Polyclonal
Conjugate
Un-conjugated
(3), (3), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Western Blotting (WB), Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
(28), (15), (9), (5), (5), (2), (1), (1), (1), (1)
Pubmed 1 reference available
Quantity 0.025 mL
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Catalog No. ABIN268449
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Immunogen A synthetic peptide made to an N-terminal region of the human xCT protein (between residues 1-50)
Purification affinity purified
Alternative Name XCT / SLC7A11
Background Mammalian amino acid transport systems consist of a large variety of transporters. Theamino acid transporter, cystine/glutamic acid transporter xCT, for system xc has beenproposed to be responsible for the cystine transport through the plasma membrane.System xc mediates an amino acid exchange and prefers cystine and glutamate as itssubstrates. The transporter designated as xCT requires 4F2 heavy chain (4F2hc:CD98)for its functional expression, and belongs to the family of amino acid transporters thatassociates with the type II membrane glycoproteins such as 4F2hc. In brain, it has beenproposed that system xc is up-regulated in glial cells upon oxidative stress and plays anessential roles to protect neurons against oxidative stress. Recent studies haveimplicated xCT as a gateway that Kaposi's sarcoma herpesvirus (KSHV) uses to entercells. Alternate Names: anti-Cysteine/glutamate transporter antibody, anti-Amino acid transport system xc-xCTantibody, anti-Calcium channel blocker resistance protein CCBR1 antibody, anti-SoluteCarrier Family 7 antibody, anti-Member 11 (SCL7a11) antibody, anti-SLC7a11 antibody.
Gene Symbol: SLC7A11
Gene ID 23657
UniProt Q9UPY5
Application Notes This xCT antibody is useful for Western blot, Immunocytochemistry/Immunofluorescence, Immunohistochemistry and Flow (PMID 20028852). Immunoprecipitation is not recommended. In Western blot this antibody recognizes a band at ~55 kDa, and in ICC/IF membrane staining was observed in HepG2 cells. Permeablization is recommended prior to performing Flow analysis.
Recommended dilutions: Flow Cytometry 2-3 µg/mL, Immunocytochemistry/Immunofluorescence 1:100-1:1000, Immunohistochemistry 5 u/gmL, Immunohistochemistry-Paraffin 5 µg/mL, Immunoprecipitation (Negative), Western Blot 1:1000
Protocol Western blot Protocol specific for xCT antibody Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
. Rinse the blot.
. Block the membrane using standard blocking buffer for at least 1 hour.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions.Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.Immunocytochemistry Protocol for NB300-318 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 1.0 mg/mL
Buffer Tris-glycine, 150 mM NaCl, 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 4 °C
Storage Comment 4 °C short term. Aliquot and store at -20 °C long term.
General Liu, Blower, Pham et al.: "Cystine-glutamate transporter SLC7A11 mediates resistance to geldanamycin but not to 17-(allylamino)-17-demethoxygeldanamycin." in: Molecular pharmacology, Vol. 72, Issue 6, pp. 1637-46, 2007 (PubMed).

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