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.

xCT Antibody (A7C6-R)

Novus Biologicals | Catalog # NBP3-33104

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Product Documents

Recombinant Monoclonal Antibody

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Product Specifications
Scientific Data
Applications
Flow Cytometry
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species Reactivity

Human, Mouse

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Mouse IgG_2B Clone # A7C6-R expressed in HEK293

View all xCT/SLC7A11 Primary Antibodies »

Product Specifications

Immunogen

293 cell line overexpressing xCT. (Uniprot: Q9UPY5)

Localization

Membrane.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2B

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 (A7C6-R)

Western Blot: xCT Antibody (A7C6-R) [NBP3-33104]

Western Blot: xCT Antibody (A7C6-R) [NBP3-33104] - Western blot analysis of xCT on different lysates with Mouse anti-xCT antibody (NBP3-33104) at 1/500 dilution.

Lane 1: HT-29 cell lysate
Lane 2: A549 cell lysate

Lysates/proteins at 10 ug/Lane.

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 30 seconds;

10% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-33104) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: xCT Antibody (A7C6-R) [NBP3-33104]

Immunohistochemistry: xCT Antibody (A7C6-R) [NBP3-33104] - Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Mouse anti-xCT antibody (NBP3-33104) at 1/600 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-33104) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: xCT Antibody (A7C6-R) [NBP3-33104]

Immunocytochemistry/ Immunofluorescence: xCT Antibody (A7C6-R) [NBP3-33104] - Immunocytochemistry analysis of HT-29 cells labeling xCT with Mouse anti-xCT antibody (NBP3-33104) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-xCT antibody (NBP3-33104) at 1/200 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Flow Cytometry: xCT Antibody (A7C6-R) [NBP3-33104]

Flow Cytometry: xCT Antibody (A7C6-R) [NBP3-33104] - Flow cytometric analysis of A549 cells labeling xCT.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (NBP3-33104, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for 30 minutes, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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Applications for xCT Antibody (A7C6-R)

Application

Recommended Usage

Flow Cytometry

1:500-1:1000

Immunocytochemistry/ Immunofluorescence

1:200

Immunohistochemistry-Paraffin

1:600

Western Blot

1:500-1:5000

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Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
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Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

Preservative

0.05% 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 (A7C6-R)

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Certificate of Analysis

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Product Specific Notices for xCT Antibody (A7C6-R)

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

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