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 (JE31-93)

Novus Biologicals | Catalog # NBP3-33014

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Product Documents

Recombinant Monoclonal Antibody

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

Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Western Blot, Immunoprecipitation

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # JE31-93 expressed in HEK293

View all xCT/SLC7A11 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide within Human xCT aa 1-50 / 501. (Uniprot: Q9UPY5)

Localization

Membrane.

Clonality

Monoclonal

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 (JE31-93)

Immunoprecipitation: xCT Antibody (JE31-93) [NBP3-33014]

Immunoprecipitation: xCT Antibody (JE31-93) [NBP3-33014] - xCT was immunoprecipitated in 0.2mg HeLa cell lysate with NBP3-33014 at 2 ug/25 ul agarose. Western blot was performed from the immunoprecipitate using NBP3-33014 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: NBP3-33014 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of NBP3-33014 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 24 seconds; ECL

Western Blot: xCT Antibody (JE31-93) [NBP3-33014]

Western Blot: xCT Antibody (JE31-93) [NBP3-33014] - Western blot analysis of xCT on different lysates with Rabbit anti-xCT antibody (NBP3-33014) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate (20 ug/Lane)
Lane 2: HeLa cell lysate (20 ug/Lane)
Lane 3: 293T cell lysate (20 ug/Lane)
Lane 4: Mouse liver tissue lysate (40 ug/Lane)
Lane 5: Mouse brain tissue lysate (40 ug/Lane)
Lane 6: Rat brain tissue lysate (40 ug/Lane)
Lane 7: Rat liver tissue lysate (40 ug/Lane)

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

Exposure time: 5 seconds;

4-20% 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 (NBP3-33014) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Applications for xCT Antibody (JE31-93)

Application

Recommended Usage

Immunoprecipitation

1-2ug/sample

Western Blot

1:1000

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

1*TBS (pH7.4), 0.05% 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 (JE31-93)

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Certificate of Analysis

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Product Specific Notices for xCT Antibody (JE31-93)

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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