xCT Antibody Pack

Dual RNAscope ISH-IHC: xCT Antibody Pack [NB100-937] - Formalin-fixed paraffin-embedded tissue sections of human stomach were probed for xCT mRNA (ACD RNAScope Probe, catalog #422688; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB300-318) at 0.25ug/mL with 1 hour incubation at room temperature followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

Simple Western: xCT Antibody Pack [NB100-937] - Simple Western lane view shows a specific band for xCT using NB300-318 at 25 ug/ml in HeLa and HeLa + DEM cell lysates. This experiment was performed under reducing conditions using the 12-230 kDa separation system. xCT Antibody [NB300-318]

Western Blot: xCT Antibody Pack [NB100-937] - Detection of XCT in total human and mouse stomach lysate, respectively, using NB 300-318. 1 minute ECL exposure.

Immunohistochemistry: xCT Antibody Pack [NB100-937] - xCT expression is enhanced in CD68+ cells from MS spinal cord. CD68+ cells (arrows) show enhanced xCT expression in MS patients as compared to controls. CD68+ macrophages are round shaped and form clusters in MS patients, whereas in controls, CD68+ cells appear isolated and long shaped. Scale bar = 50 um. Image collected and cropped by CiteAb from the following publication (//jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-8-63), licensed under a CC-BY license. xCT Antibody [NB300-317]

Simple Western: xCT Antibody Pack [NB100-937] - (1) ladder, (2) no lysate + xCT, 100ug/ml, (3) human brain frontal cortex membrane lysate,no primary antibody, (4-7) human brain lysates, 0.03mg/ml, xCT, 100ug/ml.

Western Blot: xCT Antibody Pack [NB100-937] - Total protein from human HeLa, Huvec, HCT-116 and A549 cells 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 block buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. xCT Antibody [NB300-317]

Western Blot: xCT Antibody Pack [NB100-937] - Detection of XCT in total human stomach lysate using NB 300-317. 15 minute ECL exposure.

Product Discontinued

xCT Antibody Pack Summary

• Description: This pack contains 1 vial each of: NB300-317 (0.1 mL) and NB300-318 (0.1 mL).
• Immunogen: See individual datasheets.
• Host: Rabbit
• Gene: SLC7A11

Applications/Dilutions

• Dilutions:
  • Dual RNAscope ISH-IHC
  • Immunohistochemistry
  • Simple Western
  • Western Blot
• Application Notes: See individual datasheets for validated applications.

Reactivity Notes

See individual datasheets.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Alternate Names for xCT Antibody Pack

• Amino acid transport system xc-
• Calcium channel blocker resistance protein CCBR1
• CCBR1
• cystine/glutamate transporter
• SLC7A11
• solute carrier family 7 (anionic amino acid transporter light chain, xc- system), member 11
• Solute carrier family 7 member 11
• solute carrier family 7, (cationic amino acid transporter, y+ system) member 11
• xCT

Background

xCT, also called SLC7A11, is the light chain component of the cysteine/glutamate amino acid exchange transporter system Xc (1,2). System Xc is composed of two subunits, the light chain (xCT) and the heavy chain (CD98hc, SLC3A2) and functions by cellular uptake of cysteine in exchange for glutamate in a 1:1 ratio (1,2). The human xCT gene is located on chromosome 4q28.3 and is synthesized as a 12-pass transmembrane protein with both the N- and C-terminals located intracellularly (2, 3). xCT is a 501 amino acids (aa) protein with a theoretical molecular weight of 55.4 kDa (3, 4). xCT expression serves many functional purposes in cells including redox balance, ferroptosis, and chemotherapy or cancer drug resistance (1-3, 5-7). Import of cysteine by xCT plays a role in promoting oxidative stress response as cysteine is a precursor for glutathione synthesis (2, 3, 5-7). Glutathione is a cofactor for ROS-detoxifying enzymes, including glutathione peroxidase (GPX), which help defend from cellular ROS-induced damage (2, 3, 5-7). In addition to its antioxidant role, xCT also utilizes glutathione and GPX to inhibit ferroptosis, which is iron-dependent, non-apoptotic cell-death that occurs with overproduction of lipid hydroperoxides (1-3, 5-7). As cancer cells often experience high oxidative stress, it is understandable that xCT is overexpressed in a variety of cancer types, such as acute myeloid leukemia and breast cancer, and affects cancer growth, invasion, metastasis, and prognosis (1-3, 5-7). xCT expression has also been shown to play a role in glutathione-mediated drug resistance during cancer treatment (1,5,7). However, studies have shown that xCT knockdown results in increased tumor cell death, highlighting its suitability as a druggable target (1,5,7). Specifically, the xCT inhibitors Sulfasalazine, an approved anti-inflammatory drug, and Erastin, a small molecule inhibitor, are potential therapeutic modalities for treating a variety of cancers when used in combination with radiotherapy or immunotherapy (1-3, 5-7).

References

1. Liu, J., Xia, X., & Huang, P. (2020). xCT: A Critical Molecule That Links Cancer Metabolism to Redox Signaling. Molecular therapy : the journal of the American Society of Gene Therapy. https://doi.org/10.1016/j.ymthe.2020.08.021

2. Koppula, P., Zhang, Y., Zhuang, L., & Gan, B. (2018). Amino acid transporter SLC7A11/xCT at the crossroads of regulating redox homeostasis and nutrient dependency of cancer. Cancer communications. https://doi.org/10.1186/s40880-018-0288-x

3. Lin, W., Wang, C., Liu, G., Bi, C., Wang, X., Zhou, Q., & Jin, H. (2020). SLC7A11/xCT in cancer: biological functions and therapeutic implications. American journal of cancer research.

4. xCT: Uniprot (Q9UPY5)

5. Koppula, P., Zhuang, L., & Gan, B. (2020). Cystine transporter SLC7A11/xCT in cancer: ferroptosis, nutrient dependency, and cancer therapy. Protein & cell. https://doi.org/10.1007/s13238-020-00789-5

6. Liu, L., Liu, R., Liu, Y., Li, G., Chen, Q., Liu, X., & Ma, S. (2020). Cystine-glutamate antiporter xCT as a therapeutic target for cancer. Cell biochemistry and function. https://doi.org/10.1002/cbf.3581

7. Cui, Q., Wang, J. Q., Assaraf, Y. G., Ren, L., Gupta, P., Wei, L., Ashby, C. R., Jr, Yang, D. H., & Chen, Z. S. (2018). Modulating ROS to overcome multidrug resistance in cancer. Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy. https://doi.org/10.1016/j.drup.2018.11.001

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Antibody Packs are guaranteed for 1 year from date of receipt.

Review for xCT Antibody Pack (NB100-937) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 07/30/2015

Summary

• Application: Western Blot
• Sample Tested: Human cancer cell whole cell lysate
• Species: Human

FAQs for xCT Antibody Pack (NB100-937). (Showing 1 - 1 of 1 FAQ).

1. Do you have a slc7a11 antibody that is conjugated, reacts to mouse, works with cell surface staining, does not need permeabilization?
   • SLC7A11 or xCT antibody NB300-318AF594 targets a cytoplasmic region of the protein, so it should work without permeabilization. However, the data does say it was permeabilized with saponin, so we cannot say for certain if it will work without permeabilization.

Bioinformatics

• Gene Symbol: SLC7A11