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Anti-xCT/SLC7A11 (extracellular) Antibody

Cystine/glutamate transporter, Amino acid transport system xc-, Solute carrier family 7 member 11, Calcium channel blocker resistance protein CCBR1
Cat #: ANT-111
Alternative Name Cystine/glutamate transporter, Amino acid transport system xc-, Solute carrier family 7 member 11, Calcium channel blocker resistance protein CCBR1
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)THHFKDAFSGRDTS, corresponding to amino acid residues 218-231 of mouse cystine/glutamate transporter (Accession Q9WTR6). 3rd extracellular loop.
Accession (Uniprot) Number Q9WTR6
Gene ID 26570
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat – identical; human – 11/14 amino acid residues identical.
RRID AB_2756725.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: if, ifc, ih, wb
May also work in: ic*, ip*, lci*
Western blot
  • Western blot analysis of human brain glioblastoma cell line (U-87 MG) lysate:
    Western blot analysis of human brain glioblastoma cell line (U-87 MG) lysate:
    1. Anti-xCT/SLC7A11 (extracellular) Antibody (1:200).
    2. Anti-xCT/SLC7A11 (extracellular) Antibody, preincubated with xCT/SLC7A11 (extracellular) Blocking Peptide (#BLP-NT111).
  • Western blot analysis of rat brain (lanes 1 and 4), mouse brain (lanes 2 and 5) and rat stomach (lanes 3 and 6) lysates:
    Western blot analysis of rat brain (lanes 1 and 4), mouse brain (lanes 2 and 5) and rat stomach (lanes 3 and 6) lysates:
    1-3. Anti-xCT/SLC7A11 (extracellular) Antibody (#ANT-111), (1:800).
    4-6. Anti-xCT/SLC7A11 (extracellular) Antibody, preincubated with xCT/SLC7A11 (extracellular) Blocking Peptide (#BLP-NT111).
Immunohistochemistry
  • Expression of xCT/SLC7A11 in mouse hippocampus
    Expression of xCT/SLC7A11 in mouse hippocampus
    Immunohistochemical staining of perfusion-fixed frozen mouse brain sections with Anti-xCT/SLC7A11 (extracellular) Antibody (#ANT-111), (1:300), followed by goat anti-rabbit-AlexaFluor-488. A. SLC7A11 staining in hippocampal CA1 region is detected in the pyramidal layer (P, vertical arrows) and in glial profiles in stratum radiatum (SR, horizontal arrows). B. Pre-incubation of the antibody with xCT/SLC7A11 (extracellular) Blocking Peptide (#BLP-NT111), suppresses staining. Cell nuclei are stained with DAPI (blue). P = Pyramidal layer; SR = Stratum radiatum; SO = Stratum oriens.
Indirect flow cytometry
  • Cell surface detection of xCT/SLC7A11 by indirect flow cytometry in live intact human THP-1 monocytic leukemia cells:
    Cell surface detection of xCT/SLC7A11 by indirect flow cytometry in live intact human THP-1 monocytic leukemia cells:
    ___ Cells.
    ___ Cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti-xCT/SLC7A11 (extracellular) Antibody (#ANT-111), (2.5µg) + goat-anti-rabbit-FITC.
References
  1. Verrey, F. et al. (2004) Pflugers Arch. 447, 532.
  2. Liu, R. et al. (2007) Mol. Pharmacol. 72, 1637.
Scientific background

The SLC7 family is divided into two subgroups, the cationic amino acid transporters (the CAT family, SLC7A1–4) and the glycoprotein-associated amino acid transporters (the gpaAT family, SLC7A5–11) also called light chains or catalytic chains of the hetero(di)meric amino acid transporters (HAT). The gene SLC7A11 encodes the xCT protein (also known as Cystine/glutamate transporter).

xCT, as well as other HATs, is comprised of a light chain with 12 non-glycosylated transmembrane helices and is associated with the glycosylated heavy chain 4F2hc (CD98, SLC3A2) or rBAT (SLC3A1) through a conserved disulphide bridge. The COOH-terminus of the xCT is localized intracellularly. Association of the light and heavy chains is required for surface expression of the transporter which forms the heterodimeric amino acid transport system xc−. xCT is mainly expressed macrophages, brain, retinal pigment cells, liver and kidney. xCT expression is elevated in cells requiring high glutathione synthesis, such as activated macrophages, neuronal and glial cells and other cells after glutathione depletion1.

xCT operates in a Na+-independent and electroneutral mode, exchanging extracellular anionic cystine (with pH dependence) for glutamate with a stoichiometry of 1:1. The driving force for this exchange is generated by the cystine concentration gradient (intracellular reduction) and the high intracellular concentration of glutamate.

xCT has been suggested as a mediator of treatment resistance in cancer patients. In tumor cells, xCT plays a crucial role in regulating intracellular levels of glutathione which has been broadly implicated in resistance to chemotherapy2.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 16/11/2020

Anti-xCT/SLC7A11 (extracellular) Antibody (#ANT-111) is a highly selective antibody directed against an epitope of the mouse cystine/glutamate transporter. The antibody can be used in western blot, immunohistochemistry, and live cell flow cytometry. The antibody recognizes an extracellular epitope and can potentially be used for detecting the protein in living cells. It has been designed to recognize xCT from human, mouse, and rat samples.

For research purposes only, not for human use

Applications

Specifications

Scientific Background

Citations

Citations
Western blot citations
  1. Mouse and hamster renal proximal tubular epithelial cell lysate.
    Eleftheriadis, T. et al. (2019) Biology 8, 74.
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