Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunoblotting, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all xCT/SLC7A11 Primary Antibodies »

Product Specifications

Immunogen

This xCT Antibody was prepared from a synthetic peptide made to a region within the N-terminus of the mouse xCT protein (between residues 1-50). [Uniprot: Q9WTR6]

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: orangutan (82%). Mouse reactivity reported in scientific literature (PMID: 30279737).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

55 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for xCT Antibody - BSA Free

Western Blot: xCT AntibodyBSA Free [NB300-317]

Western Blot: xCT AntibodyBSA Free [NB300-317]

Western Blot: xCT Antibody [NB300-317] - 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.
Immunohistochemistry-Paraffin: xCT Antibody - BSA Free [NB300-317]

Immunohistochemistry-Paraffin: xCT Antibody - BSA Free [NB300-317]

xCT-Antibody-Immunohistochemistry-Paraffin-NB300-317-img0015.jpg
Western Blot: xCT AntibodyBSA Free [NB300-317]

Western Blot: xCT AntibodyBSA Free [NB300-317]

Western Blot: xCT Antibody [NB300-317] - 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.
Flow Cytometry: xCT Antibody - BSA Free [NB300-317]

Flow Cytometry: xCT Antibody - BSA Free [NB300-317]

Flow Cytometry: xCT Antibody [NB300-317] - An intracellular stain was performed on HeLa cells with NB300-317G (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 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 488.
Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317]

Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317]

Immunocytochemistry/Immunofluorescence: xCT Antibody [NB300-317] - xCT antibody was tested in HepG2 cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).
Flow Cytometry: xCT Antibody - BSA Free [NB300-317]

Flow Cytometry: xCT Antibody - BSA Free [NB300-317]

Flow Cytometry: xCT Antibody [NB300-317] - An intracellular stain was performed on HeLa with NB300-317 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).
Simple Western: xCT AntibodyBSA Free [NB300-317]

Simple Western: xCT AntibodyBSA Free [NB300-317]

Simple Western: xCT Antibody [NB300-317] - (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.
xCT Antibody - BSA Free Immunohistochemistry-Paraffin: xCT Antibody [NB300-317]

Immunohistochemistry-Paraffin: xCT Antibody [NB300-317]

Analysis of a FFPE tissue section of human stomach using 1:200 dilution of xCT antibody. The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.
xCT Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317] -

Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317] - xCT expression is enhanced in CD68+ cells from MS spinal cord. A. Triple immunofluorescence staining for xCT (green), CD68 (red) & Hoechst 33258 (blue) in spinal cord of control (left) & MS patients (right). A high expression of xCT was detected in CD68+ infiltrating macrophages (arrows) associated with blood vessels, which are virtually absent in controls. Note that overall xCT expression is enhanced in MS tissue. B. CD68+ cells (arrows) show enhanced xCT expression in MS patients as compared to controls. CD68+ macrophages are round shaped & form clusters in MS patients, whereas in controls, CD68+ cells appear isolated & long shaped. Scale bar = 50 μm. Image collected & cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
xCT Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317] -

Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317] - xCT expression is enhanced in CD68+ cells from MS spinal cord. A. Triple immunofluorescence staining for xCT (green), CD68 (red) & Hoechst 33258 (blue) in spinal cord of control (left) & MS patients (right). A high expression of xCT was detected in CD68+ infiltrating macrophages (arrows) associated with blood vessels, which are virtually absent in controls. Note that overall xCT expression is enhanced in MS tissue. B. CD68+ cells (arrows) show enhanced xCT expression in MS patients as compared to controls. CD68+ macrophages are round shaped & form clusters in MS patients, whereas in controls, CD68+ cells appear isolated & long shaped. Scale bar = 50 μm. Image collected & cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
xCT Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317] -

Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317] - xCT expression is increased in the CNS of rats with EAE. A. Histogram showing the neurological score during the course of acute EAE induced in Lewis rats by immunization with myelin basic protein. The peak of neurological disability was at day 14 post-immunization, which was selected for obtaining tissue samples. B. xCT mRNA (left) & protein (right) expression in spinal cord from control & acute EAE rats, as assessed by qPCR & Western blot analysis. Data are referred to mean expression level of controls (n = 5-6). C. Double immunofluorescence for xCT (green) & OX-42 (red), a marker of microglia & infiltrating macrophages. OX42+ cells express high xCT levels in acute EAE. Both meninges (asterisk in top) & infiltrating cells (bottom) in inflammatory foci show high levels of xCT in rat spinal cord with EAE as compared to controls. D. Microglial cells (OX42+ cells) of EAE rats have higher xCT levels in spinal cord than controls. Notice the difference between resting microglia in control rats, with ramified morphology (arrows in control) & microglia in EAE showing round shaped morphology, characteristic of its activated state (arrows in EAE). Scale bar = 20 μm.E. xCT mRNA (left) & protein (right) expression in spinal cord from control & chronic EAE mice, assessed by qPCR & Western blot analysis. Data are referred to mean expression level of controls (n = 5). Image collected & cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
xCT Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317] -

Immunocytochemistry/ Immunofluorescence: xCT Antibody - BSA Free [NB300-317] - xCT expression is increased in the CNS of rats with EAE. A. Histogram showing the neurological score during the course of acute EAE induced in Lewis rats by immunization with myelin basic protein. The peak of neurological disability was at day 14 post-immunization, which was selected for obtaining tissue samples. B. xCT mRNA (left) & protein (right) expression in spinal cord from control & acute EAE rats, as assessed by qPCR & Western blot analysis. Data are referred to mean expression level of controls (n = 5-6). C. Double immunofluorescence for xCT (green) & OX-42 (red), a marker of microglia & infiltrating macrophages. OX42+ cells express high xCT levels in acute EAE. Both meninges (asterisk in top) & infiltrating cells (bottom) in inflammatory foci show high levels of xCT in rat spinal cord with EAE as compared to controls. D. Microglial cells (OX42+ cells) of EAE rats have higher xCT levels in spinal cord than controls. Notice the difference between resting microglia in control rats, with ramified morphology (arrows in control) & microglia in EAE showing round shaped morphology, characteristic of its activated state (arrows in EAE). Scale bar = 20 μm.E. xCT mRNA (left) & protein (right) expression in spinal cord from control & chronic EAE mice, assessed by qPCR & Western blot analysis. Data are referred to mean expression level of controls (n = 5). Image collected & cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for xCT Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

1 - 5 ug/ml

Immunoblotting

reported in scientific literature (PMID 28185919)

Immunocytochemistry/ Immunofluorescence

1:100 - 1:1000

Immunohistochemistry

1:200

Immunohistochemistry-Frozen

1:10-1:500. Use reported in scientific literature (PMID 21639880)

Immunohistochemistry-Paraffin

1:200

Western Blot

0.5-2 ug/ml
Application Notes
In Western blot, a band is observed at ~35 kDa. In ICC/IF, membrane and ER staining was visualized in HepG2 cells.

Reviewed Applications

Read 1 review rated 4 using NB300-317 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

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

Background: xCT/SLC7A11

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

Long Name

Cationic 1/Solute Carrier Family 7 Member 11

Alternate Names

CCBR1, SLC7A11

Gene Symbol

SLC7A11

Additional xCT/SLC7A11 Products

Product Documents for xCT Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for xCT Antibody - BSA Free

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

Related Research Areas

Infectious Diseases

Citations for xCT Antibody - BSA Free

Customer Reviews for xCT Antibody - BSA Free (1)

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  • xCT Antibody
    Name: Diane Brooks
    Application: Simple Western
    Sample Tested: Human Brain Tissue and human glioblastoma
    Species: Human
    Verified Customer | Posted 01/18/2018
    1:ladder, 2:no lysate + xCT (100µg/ml), 3:human brain frontal cortex membrane lysate,no primary antibody, 4:SNB-19 cell line lysate, 0.1mg/ml +xCT 20µg/ml, 5-8:human brain lysates, 0.03mg/ml, xCT, 100µg/ml.
    Membrane lysates were extracted using ThermoFisher Mem-PER Plus Membrane Protein Extraction Kit Cat# 89842. Samples were reduced following Simple Western kit instructions in 40mM DTT for 5 minutes at 95oC and separated using the 12-230kDa separation kit. Primary antibody followed by HRP-conjugated Anti-Rabbit IgG secondary antibody (Protein Simple Cat # 042-206).
    xCT Antibody - BSA Free NB300-317

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Protocols

View specific protocols for xCT Antibody - BSA Free (NB300-317):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

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FAQs for xCT Antibody - BSA Free

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  • Q: Do you have a slc7a11 antibody that is conjugated, reacts to mouse, works with cell surface staining, does not need permeabilization?

    A: 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.

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