xCT Antibody

Images

 
Orthogonal Strategies: Dual RNAscope ISH-IHC: xCT Antibody [NB300-318] - Formalin-fixed paraffin-embedded tissue sections of human stomach were probed for xCT mRNA (ACD RNAScope Probe, catalog #422688; Fast Red ...read more
Biological Strategies: Simple Western: xCT Antibody [NB300-318] - 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 ...read more
Western Blot: xCT Antibody [NB300-318] - Western blot analysis of protein isolated from subcutaneous tumors derived from in vivo growth of the clones relative to WT-derived tumors revealed that xCT levels remained low, ...read more
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 NB300-318AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with ...read more
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).
Flow Cytometry: xCT Antibody [NB300-318] - An intracellular stain was performed on HeLa cells with xCT Antibody NB300-318AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then ...read more
Biological Strategies: 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 ...read more
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. ...read more
Simple Western: xCT Antibody [NB300-318] - Simple Western lane view shows a specific band for xCT using NB300-318 at 25 ug/mL in HepG2 cell lysates and antibody at 25 ug/mL. Electropherogram image of corresponding ...read more

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, Dual ISH-IHC, IP (-)
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml

xCT Antibody Summary

Immunogen
This xCT Antibody was prepared from a synthetic peptide made to an N-terminal region of the human xCT protein (between residues 1-50) [UniProt Q9UPY5].
Localization
Membrane
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
SLC7A11
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:1000
  • Simple Western 10 ug/ml
  • Flow Cytometry 2-3ug/ml
  • Immunocytochemistry/Immunofluorescence 1:100-1:1000
  • Immunohistochemistry 5 u/gml
  • Immunohistochemistry-Paraffin 5 ug/ml
  • Dual RNAscope ISH-IHC
Application Notes
This xCT antibody is useful for Western blot, Immunocytochemistry/Immunofluorescence, Immunohistochemistry and Flow (PMID: 20028852). Immunoprecipitation is not recommended. In Western blot this antibody recognizes a band at approx. 35 kDa, and in ICC/IF membrane staining was observed in HeLa cells. Permeablization is recommended prior to performing Flow analysis.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.
Control
xCT Overexpression Lysate
Human Stomach Whole Tissue Lysate (Adult Whole Normal)
Control Peptide
xCT Protein (NB300-318PEP)
Reviewed Applications
Read 3 Reviews rated 4.3
using
NB300-318 in the following applications:

Publications
Read Publications using
NB300-318 in the following applications:

  • 5 publications
  • IHC
    5 publications
  • 3 publications
  • WB
    33 publications

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 21540084).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for xCT Antibody

  • 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. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for xCT Antibody (NB300-318)(56)

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 4 applications: ICC/IF, IHC, IHC-P, WB.


Filter By Application
ICC/IF
(5)
IHC
(5)
IHC-P
(3)
WB
(33)
All Applications
Filter By Species
Human
(21)
Mouse
(12)
Rat
(8)
All Species
Showing Publications 1 - 10 of 56. Show All 56 Publications.
Publications using NB300-318 Applications Species
Kazama M, Kato Y, Kakita A et al. Astrocytes release glutamate via cystine/glutamate antiporter upregulated in response to increased oxidative stress related to sporadic amyotrophic lateral sclerosis Neuropathology : official journal of the Japanese Society of Neuropathology Dec 1 2020 [PMID: 33305472] (IHC) IHC
Healey KL, Kibble S, Bell A et al. Effects of adolescent intermittent ethanol on hippocampal expression of glutamate homeostasis and astrocyte-neuronal tethering proteins in male and female rats Cell Metab Nov 13 2020 [PMID: 33217775]
Krys D, Mattingly S, Glubrecht D et al. PET Imaging of L-Type Amino Acid Transporter (LAT1) and Cystine-Glutamate Antiporter (xc-) with [18F]FDOPA and [18F]FSPG in Breast Cancer Models Mol Imaging Biol Aug 13 2020 [PMID: 32789819]
Park SY, Na SJ, Kumar M et al. Clinical evaluation of (4S)-4-(3-[18F]Fluoropropyl)-L-glutamate (18F-FSPG) for PET/CT imaging in patients with newly diagnosed and recurrent prostate cancer Clin. Cancer Res. Jul 21 2020 [PMID: 32694158] (IHC-P, IHC, Human) IHC-P, IHC Human
Gao M, Liu H, Xiao Y et al. xCT regulates redox homeostasis and promotes photoreceptor survival after retinal detachment Free Radic. Biol. Med. Jul 15 2020 [PMID: 32679366] (WB, ICC/IF, Mouse) WB, ICC/IF Mouse
Long Y, Tao H, Karachi A et al. Dysregulation of glutamate transport enhances Treg function that promotes VEGF blockade resistance in glioblastoma Cancer Res. Nov 13 2019 [PMID: 31723000]
Wu WC, Sun HW, Chen J et al. Immunosuppressive immature myeloid cell generation is controlled by glutamine metabolism in human cancer Cancer Immunol Res Aug 6 2019 [PMID: 31387898] (IHC-P, ICC/IF, Mouse) IHC-P, ICC/IF Mouse
Pitman KE, Alluri SR, Kristian A et al. Influx rate of 18F-fluoroaminosuberic acid reflects cystine/glutamate antiporter expression in tumour xenografts Eur. J. Nucl. Med. Mol. Imaging Sep 1 2019 [PMID: 31264167] (WB, Mouse) WB Mouse
Isobe S, Kataoka M, Endo J et al. Endothelial-Mesenchymal Transition Drives Expression of CD44 Variant and xCT in Pulmonary Hypertension Am. J. Respir. Cell Mol. Biol. Mar 21 2019 [PMID: 30897333] (ICC/IF, Human) ICC/IF Human
Ungard R, Linher-Melville K, Nashed M et al. xCT knockdown in human breast cancer cells delays onset of cancer-induced bone pain Mol Pain Jan 1 2019 [PMID: 30799686] (WB, Human) WB Human
Show All 56 Publications.

Reviews for xCT Antibody (NB300-318) (3) 4.33

Average Rating: 4.3
(Based on 3 reviews)
We have 3 reviews tested in 2 species: Human, Mouse.

Reviews using NB300-318:
Filter by Applications
WB
(3)
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Human
(2)
Mouse
(1)
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Images Ratings Applications Species Date Details
Western Blot xCT NB300-318
Enlarge
4
reviewed by:
Anonymous
WB Human 09/18/2018
View

Summary

ApplicationWestern Blot
Sample TestedHuman Cancer Cells
SpeciesHuman
LotG-2
Western Blot xCT NB300-318
Enlarge
5
reviewed by:
Dan Cojocari
WB Human 12/24/2015
View

Summary

ApplicationWestern Blot
Sample Testedpancreatic cancer cell line lysate
SpeciesHuman
LotH1

Blocking

Blocking DetailsLicor Odyssey Blocking buffer, 1 hour at room temperature

Primary Anitbody

Dilution Ratio1000x dilution, overnight incubation at 4C in 0.1% Twin-20 in TBS

Secondary Antibody

Secondary DescriptionLi-cor IRDye® 800CW Donkey anti-Rabbit IgG (H+L)
Secondary Manufacturer Cat#LIC-926-32213
Secondary Concentration10,000 dilution

Details

Detection NotesLi-cor Odyssey CLx
Western Blot xCT NB300-318
Enlarge
4
reviewed by:
Anonymous
WB Mouse 05/01/2013
View

Summary

ApplicationWestern Blot
Sample TestedMouse Whole Brain
SpeciesMouse
LotF-4
CommentsBiorad Bradford Assay was done to make sure that 10mcg of protein were used for each mouse. I thought the antibody was quite easy to use.

Blocking

Blocking Details5% Milk in PBTX, 1 hr

Primary Anitbody

Dilution Ratio1:1000, overnight 4 degrees C, in 5% milk/PBTX

Secondary Antibody

Secondary DescriptionAnti-rabbit
Secondary Concentration1:100,000

Details

Detection Notes5 min Bio-rad Clarity ECL, film exposure for 30 minutes, three negative controls, xCT knockouts

Comments

CommentsBiorad Bradford Assay was done to make sure that 10mcg of protein were used for each mouse. I thought the antibody was quite easy to use.

Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for xCT Antibody (NB300-318). (Showing 1 - 10 of 10 FAQs).

  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.
  2. I am using the xCT antibody, NB300-318, for IHC. I was wondering if you could recommend a rabbit negative isotype control IGg/serum?
    • I would recommend catalog number NB810-56910 as a rabbit isotype control for NB300-318.
  3. I am using the xCT antibody, NB300-318, for IHC. I was wondering if you could recommend a peptide to use for a negative control?
    • I regret to inform you that we do not have a blocking peptide available for this antibody at this time.
  4. Is there a blocking peptide available for use with NB300-318?
    • We unfortunately do not carry the blocking peptide to NB300-318.
  5. We ordered some xCT antibody (NB300-318). Is itpossible to know if there is -NH2 residues on this synthetic peptide. If so, how many -NH2 or lysine are there?
    • The immunogen for NB300-318 contains 1 lysine residue.
  6. For xCT antibody [NB300-318], why do you see a band at 35kDa when the predicted MW is ~55kDa and xCT is not known to undergo cleavage?
    • Although the theoretical molecular weight of xCT is ~55kDa, the protein’s amino acid structure, with multiple charged residues, results in its fast migration. The research paper by Liefferinge et al 2016 (The Journal of Comparative Neurology 524:1015-1032) shows the testing of several different xCT antibodies in knockout study and it was confirmed that the protein indeed runs at 35kDa (Fig.2. lane 1&2 lanes are xCT positive and lanes 3&4 are from xCT knockouts).  
  7. Hello, I'm trying to run a western blot with NB300-318. What is the blocking buffer used?
    • For NB300-318 we recommend using 5% milk to block Blocking solution 1X TBST 5% non-fat dry milk
  8. Is NB300-318 suitable for live cell flow cytometry? (is its epitope in the external side of the cells?) I would need it for FACS sorting application
    • The immunogen for this antibody is cytoplamic, so this antibody would not be suitable for live flow.
  9. I am using the xCT antibody NB300-318 for IHC.  I was wondering if you could recommend  a rabbit negative isotype control IGg/serum?
    •  I would recommend this isotype control for you: www.novusbio.com/Rabbit-IgG-Isotype-Control_NB810-56910.html
  10. We are wondering if it is possible to know if there is -NH2 residues on this synthetic peptide. If yes, how many -NH2 or lysine are there?
    • The immunogen for NB300-318 contains 1 lysine residue.  

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB300-318AF350
Alexa Fluor 405 NB300-318AF405
Alexa Fluor 488 NB300-318AF488
Alexa Fluor 532 NB300-318AF532
Alexa Fluor 594 NB300-318AF594
Alexa Fluor 647 NB300-318AF647
Alexa Fluor 700 NB300-318AF700
Alexa Fluor 750 NB300-318AF750
Biotin NB300-318B
DyLight 350 NB300-318UV
DyLight 405 NB300-318V
DyLight 488 NB300-318G
DyLight 550 NB300-318R
DyLight 594 NB300-318DL594
DyLight 650 NB300-318C
DyLight 680 NB300-318FR
DyLight 755 NB300-318IR
FITC NB300-318F
HRP NB300-318H
Janelia Fluor 549 NB300-318JF549
Janelia Fluor 646 NB300-318JF646
PE NB300-318PE

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Bioinformatics Tool for xCT Antibody (NB300-318)

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Blogs on xCT.

The effects of ethanol consumption on glutamate production and xCT
xCT is a sodium independent glutamate transporter that regulates the exchange of extracellular l-cystine and intracellular l-glutamate across the plasma membrane. This process is critical to glutathione production and protection from subsequent ox...  Read full blog post.

xCT: The Membrane's Gatekeeper
xCT is an obligate, electroneutral, membrane-bound anionic transporter responsible for regulating particular amino acid gradients via their transport through plasma membrane. The antiporter xCT superficially resembles an ion channel and preferentially...  Read full blog post.

xCT: Amino Acid Transport and Disorders of the Central Nervous System
xCT, encoded by the gene SLC7A11, is a member of the heterodimeric amino acid transporter family. Proteins within this family are linked to one another via a disulphide bond to form heterodimers consisting of one light subunit and one heavy subunit (1...  Read full blog post.

xCT: Friend or Foe?
There are two opposing sides to the controversial cysteine/glutamate antiporter. On one hand, it can be viewed a guardian of the cell, protecting it from the damaging oxidative stress that can cause cell death and even cancer. But, conversely, it has...  Read full blog post.

Glutathione and xCT: Chemoresistance in Tumor Cells
Glutathione, called GSH in its reduced form and GSSG or L(-)-Glutathione in its oxidized form, is an endogenous antioxidant found in most cells in the body. Glutathione's functions include detoxifying xenobiotics from the body, assisting in membrane t...  Read full blog post.

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Recent Reviews

4.3
5
1
4
2
3
0
2
0
1
0

Anonymous
09/18/2018
Application: WB
Species: Human

Dan Cojocari
12/24/2015
Application: WB
Species: Human

Anonymous
05/01/2013
Application: WB
Species: Mouse

Bioinformatics

Gene Symbol SLC7A11
Entrez
Uniprot