抗xCT抗体 | Anti-xCT Antibody

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	Biological Strategies Validation. Western Blot: xCT Antibody [NB300-318] - Total protein from Human HeLa cells treated with and without 0.1 mM Diethyl Maleate for 24 hours was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-xCT in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the increase in xCT expression with treatment.
	Immunocytochemistry/Immunofluorescence: xCT Antibody [NB300-318] - xCT antibody was tested in HepG2 cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
	Immunohistochemistry: xCT Antibody [NB300-318] - xCT staining in the absorptive epithelia of small intestinal villi detected using NB300-318.
	Flow Cytometry: xCT Antibody [NB300-318] - An intracellular stain was performed on HeLa cells with xCT Antibody NB300-318AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.
	Flow Cytometry: xCT Antibody [NB300-318] - An intracellular stain was performed on HeLa with xCT Antibody NB300-318 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG APC-conjugated Secondary Antibody (F0111, R&D Systems).

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Background

The amino acid transporter, cystine/glutamic acid transporter xCT, for system xc has been proposed to be responsible for the cystine transport through the plasma membrane. System xc mediates an amino acid exchange and prefers cystine and glutamate as its substrates. The transporter designated as xCT requires 4F2 heavy chain (4F2hc:CD98) for its functional expression, and belongs to the family of amino acid transporters that associates with the type II membrane glycoproteins such as 4F2hc. In brain, it has been proposed that system xc is up-regulated in glial cells upon oxidative stress and plays an essential roles to protect neurons against oxidative stress. Recent studies have implicated xCT as a gateway that Kaposi's sarcoma herpesvirus (KSHV) uses to enter cells.

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