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  1. Link

    products/primary-antibodies/xct-antibody-ab37185.pdf

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Signal Transduction Metabolism Amino Acids
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Anti-xCT antibody (ab37185)

  • Datasheet
  • SDS
Reviews (2)Q&A (8)References (87)

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Immunocytochemistry/ Immunofluorescence - Anti-xCT antibody (ab37185)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-xCT antibody (ab37185)

Key features and details

  • Rabbit polyclonal to xCT
  • Suitable for: IHC-P, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-xCT antibody [EPR27115-64] (ab307601)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-xCT antibody
    See all xCT primary antibodies
  • Description

    Rabbit polyclonal to xCT
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human xCT aa 1-50 (N terminal). The exact sequence is proprietary.
    Sequence:

    MVRKPVVSTISKGGYLQGNVNGRLPSLGNKEPPGQEKVQLKRKVTLLRGV


    Database link: Q9UPY5
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • ICC/IF: HepG2 cells. IHC-P: Human small intestine tissue.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    Affinity purified antibody.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Amino Acids
    • Signal Transduction
    • Metabolism
    • Plasma Membrane
    • Channels
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Amino acid metabolism
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human xCT protein (ab112403)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab37185 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P
Use at an assay dependent concentration.
ICC/IF
1/100 - 1/1000.
Notes
IHC-P
Use at an assay dependent concentration.
ICC/IF
1/100 - 1/1000.

Target

  • Function

    Sodium-independent, high-affinity exchange of anionic amino acids with high specificity for anionic form of cystine and glutamate.
  • Sequence similarities

    Belongs to the amino acid-polyamine-organocation (APC) superfamily. L-type amino acid transporter (LAT) (TC 2.A.3.8) family.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession Q9UPY5 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 23657 Human
    • Omim: 607933 Human
    • SwissProt: Q9UPY5 Human
    • Unigene: 390594 Human
    • Form

      xCT has a predicted molecular weight of 55 kDa; however it has a high number of hydrophobic residues which may affect the migration of the protein in SDS-PAGE. Endogenous monomeric xCT is expected to migrate at ~35 kDa and modified-xCT is expected to migrate at ~55 kDa (PMID: 17035536)
    • Alternative names

      • Amino acid transport system xc xCT antibody antibody
      • Amino acid transport system xc- antibody
      • Calcium channel blocker resistance protein CCBR1 antibody
      • Calcium channel blocker resistance protein CCBR1 antibody antibody
      • CCBR1 antibody
      • Cysteine/glutamate transporter antibody antibody
      • Cystine/glutamate transporter antibody
      • OTTHUMP00000164578 antibody
      • SLC7A11 antibody
      • Solute carrier family 7 (anionic amino acid transporter light chain, xc- system), member 11 antibody
      • solute carrier family 7 antibody
      • Solute carrier family 7 member 11 antibody
      • Solute carrier family 7, (cationic amino acid transporter, y+ system) member 11 antibody
      • SYSTEM Xc(-) TRANSPORTER-RELATED PROTEIN antibody
      • xCT antibody
      • XCT_HUMAN antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-xCT antibody (ab37185)
      Immunocytochemistry/ Immunofluorescence - Anti-xCT antibody (ab37185)

      Imunocytochemistry/ Immunofluorescence analysis of HepG2 cells labeling xCT with ab37185 followed by Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red) respectively. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-xCT antibody (ab37185)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-xCT antibody (ab37185)

      Immunohistochemical analysis of human small intestine labeling xCT with ab37185. Staining is observed in the absorptive epithelia of small intestinal villi. 

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (87)

    Publishing research using ab37185? Please let us know so that we can cite the reference in this datasheet.

    ab37185 has been referenced in 87 publications.

    • Müller F  et al. Elevated FSP1 protects KRAS-mutated cells from ferroptosis during tumor initiation. Cell Death Differ 30:442-456 (2023). PubMed: 36443441
    • Wei TT  et al. Interferon-γ induces retinal pigment epithelial cell Ferroptosis by a JAK1-2/STAT1/SLC7A11 signaling pathway in Age-related Macular Degeneration. FEBS J 289:1968-1983 (2022). PubMed: 34741776
    • Sendo K  et al. Impact of the glutathione synthesis pathway on sulfasalazine-treated endometrial cancer. Oncotarget 13:224-236 (2022). PubMed: 35106124
    • Huang S  et al. Hepatic TGFβr1 Deficiency Attenuates Lipopolysaccharide/D-Galactosamine-Induced Acute Liver Failure Through Inhibiting GSK3β-Nrf2-Mediated Hepatocyte Apoptosis and Ferroptosis. Cell Mol Gastroenterol Hepatol 13:1649-1672 (2022). PubMed: 35202887
    • Zhang R  et al. Curcumenol triggered ferroptosis in lung cancer cells via lncRNA H19/miR-19b-3p/FTH1 axis. Bioact Mater 13:23-36 (2022). PubMed: 35224289
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 10 Abreviews or Q&A

    Western blot abreview for Anti-xCT antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (head and neck cancer)
    Gel Running Conditions
    Non-reduced Denaturing (10% gel)
    Loading amount
    20 µg
    Treatment
    drug 48h
    Specification
    head and neck cancer
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 21 2015

    IHC - Wholemount abreview for Anti-xCT antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Human Tissue (head and neck cancer)
    Specification
    head and neck cancer
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Miss. Jin Young Park

    Verified customer

    Submitted Oct 19 2015

    Question



    You ask a good question. Actually, A549 cells have been shown to have high expression of xCT, so that is my positive control. After doing a bit of research on the matter, I think that the 105 kDa band that I am detecting is probably the heterodimer of xCT with 4F2hc and that the 35 kDa band is xCT. I did use B-mercaptoethanol to reduce disulfides, but the reaction may have been inefficient due to the age of the B-me. I am still unsure of the identity of the other bands, though. In my next study, I am going to analyze the samples +/- B-me (fresh solution) to see if the 105 kDa band reduces to the 35 kDa form.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 10 2012

    Answer

    Thanks so muchfor your reply and best of luck with your upcoming experiments. If you have any further questions please do not hesitate to contact me.

    Read More

    Abcam Scientific Support

    Answered on Jul 10 2012

    Question

    Inquiry: Hello, I have made two attempts to use your anti-xCT antibody to detect xCT in cell lysates and both blots have resulted in numerous bands, none at the predicted MW of 55 kDa. I have attached an image of my most recent blot to this message. My method consists of loading whole cell lysate (25 and 10 ug), blocking in 5% milk/TBS-T 1 h at RT, xCT antibody incubation at 0.5 ug/mL overnight at 4 deg C in 5% milk/TBS-T, anti-rabbit IgG-HRP (Rockland) at 1:5000 1 h at RT in 5% milk/TBS-T, and chemiluminescent detection with Supersignal Pico (Pierce). I would appreciate any advice that could be given for modifying my conditions or any insight into the identity of the nonspecific bands.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 09 2012

    Answer

    Thanks for submitting your enquiry. I am sorry to hear this antibody is giving you trouble.

    I was just wondering what positive control samples you have used?

    Thanks in advance for the additional information.

    Read More

    Abcam Scientific Support

    Answered on Jul 09 2012

    Question

    Hi There,

     

    I wrote you emails to explain my concern of using your xCT antibody to do flow cytometry and ask for your help. But I did not receive any response after I provided you with my experiment details, which makes me very upset. My lab ordered this specific antibody from your company at least 4 times, I think I deserve better service!

    Read More

    Abcam community

    Verified customer

    Asked on Apr 27 2012

    Answer

    Thank you for contacting us and making us aware of this. We have been transitioning to a new email system and it may be that the response which I had sent on Mon 16 Apr 12 13:26GMT was somehow caught up or perhaps it went directly to your spam or junk mail boxes. Regardless prompt customer service is very important to us andI am sorry if it appears that your response was not sent in a timely manner to you at XXXX. If you could please send me the order number of the xCT antibody which you have purchased I would be happy to use that information to create a token of our appreciation. I have pasted the body the email response here: Thank you for your response and for the additional details which you have provided. You are correct as this antibody will only recognize the intracellular epitope of xCT. As such you would need to permeablize your cells to allow entry of the antibody. In the publication:Pampliega Oet al.Increased expression of cystine/glutamate antiporter in multiple sclerosis.J Neuroinflammation8:63 (2011).http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=21639880&dopt=Abstractthey were able to use this product, conjugated to FITC with a concentration of 2.1ug/ml. Of course the appropriate concentrations would need to be determined experimentally in your protocol but this is an excellent starting point. At Abcam we do offer a number of different antibody conjugation kits for fast, easy and efficient direct conjugation of chromogen or enzymatic or biotin/streptavidin tag to your antibodies. We do recommend direct conjugation of the antibody in Flow cytometry as this eliminates the need for secondary antibodies and reduces complexity of the protocol. More information about our EasyLink direct conjugation kits may be found at the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=13148 Between April 1rst and May 31rst we are running a special promotion where you may receive a FREE primary antibody with purchase of an EasyLink conjugation kit. If you are interested in this offer please follow the link below: https://www.abcam.com/index.html?pageconfig=resource&rid=14962 Alternately, I would recommend using Ab111822 as this product has been produced from an extracellular region of xCT,amino acids 215-229. As this product has not been tested in Flow Cytometry however we cannot guarantee its effectiveness in that application. However we may offer to you our 100% Abreview testing discount should you wish to use this in Flow. With this program, you would purchase Ab111822, test in Flow Cytometry and leave an "Abreview". After the Abreview has been submitted, a code which I will provide to you will become active, entitling you to on e free primary antibody of your choice. If you are interested in participating in our 100% Abreview testing discount program please let me know and I will send further information to you. I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

    Read More

    Abcam Scientific Support

    Answered on Apr 27 2012

    Question

    I used human breast cancer cell line MCF-7 cells. Negative control: cells only, secondary anti rabbit IgG fitc only. No positive control. I used up the antibody for real experiments.. FITC secondary antibody was newly ordered, it is working very well in our lab's experiments. I did not try to directly conjugated a tag to the antibody. How to do that? 100000 cells per sample in 500ul flow buffer (BSA + PBS + N3NA). I put 10ul of you shipped antibody per test. I tried 5 ul to 10 ul of the antibody. Never works. (what's your recommended concentration? I did not find the information in the data sheet..) My cells should be live cells. I did not use live/dead dye.   Your antibody targets the N terminal of xCT protein. I searched xCT's structure, N terminal locates intracellularly. Does that mean I could not use the antibody to target cell surface expression of xCT protein? It seems only your company carry xCT ab for flow. I am quite desperate to get it to work! Thanks!  

    Read More

    Abcam community

    Verified customer

    Asked on Apr 16 2012

    Answer

    Thank you for your response and for the additional details which you have provided.

    You are correct as this antibody will only recognize the intracellular epitope of xCT. As such you would need to permeablize your cells to allow entry of the antibody. In the publication:Pampliega Oet al.Increased expression of cystine/glutamate antiporter in multiple sclerosis.J Neuroinflammation8:63 (2011).http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=21639880&dopt=Abstractthey were able to use this product, conjugated to FITC with a concentration of 2.1ug/ml. Of course the appropriate concentrations would need to be determined experimentally in your protocol but this is an excellent starting point.

    At Abcam we do offer a number of different antibody conjugation kits for fast, easy and efficient direct conjugation of chromogen or enzymatic or biotin/streptavidin tag to your antibodies. We do recommend direct conjugation of the antibody in Flow cytometry as this eliminates the need for secondary antibodies and reduces complexity of the protocol. More information about our EasyLink direct conjugation kits may be found at the following link:

    https://www.abcam.com/index.html?pageconfig=resource&rid=13148

    Between April 1rst and May 31rst we are running a special promotion where you may receive a FREE primary antibody with purchase of an EasyLink conjugation kit. If you are interested in this offer please follow the link below:

    https://www.abcam.com/index.html?pageconfig=resource&rid=14962

    Alternately, I would recommend using Ab111822 as this product has been produced from an extracellular region of xCT,amino acids 215-229. As this product has not been tested in Flow Cytometry however we cannot guarantee its effectiveness in that application. However we may offer to you our 100% Abreview testing discount should you wish to use this in Flow. With this program, you would purchase Ab111822, test in Flow Cytometry and leave an "Abreview". After the Abreview has been submitted, a code which I will provide to you will become active, entitling you to on e free primary antibody of your choice. If you are interested in participating in our 100% Abreview testing discount program please let me know and I will send further information to you.

    I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

    Read More

    Abcam Scientific Support

    Answered on Apr 16 2012

    Question

    Hi There,

    Our lab recently purchased xCT antibody ab37185 for flow cytometry study. I want to measure xCT cell surface expression. So I first incubated my cells with ab37185 (10ul per sample) for 1hr and washed for 2˜3 times with PBS then incubated the cells with anti-rabbit IgG FITC antibody. However, I could never get a signal by flow. I am wondering what's wrong. Can you give me some suggestions? Thanks!

    Read More

    Abcam community

    Verified customer

    Asked on Apr 13 2012

    Answer

    Thank you for contacting Abcam, I am sorry to hear that you have been experiencing difficulties using this product in flow.

    Would you be able to provide me the specific details of your experiments? Useful information would include but not be limited to:

    Cell type, species and any treatments. Isotype controls used. Positive controls.
    Have you used the secondary antibody with luck in flow?
    Have you attempted to directly conjugate a tag to the antibody?
    How many cells were used per experiment? What was the concentration of your lot and overall concentration of the antibody in your cell solution?
    Have you attempted different concentrations?
    Did you use a live/dead dye?
    Any data is also very helpful.

    I would also suggest reviewingPampliega O et al.Increased expression of cystine/glutamate antiporter in multiple sclerosis.J Neuroinflammation8:63 (2011). WB, Flow Cyt, IHC-FoFr; Human, Mouse, Rat.http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=21639880&dopt=Abstract, as they did extensive testing with this product in flow cytometry.

    This product is covered by our Abpromise guarantee. If we cannot remedy this issue and this is a product that you have purchased within the last six months, we will replace or refund it under our Abpromise guarantee, as you are using it according to specifications listed on our datasheet.

    I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

    Read More

    Abcam Scientific Support

    Answered on Apr 14 2012

    Question

    I'm wondering if you could send me a aliquot of your xCT antibody ab37185.
    Indeed, I would like to test it on fresh samplesthat will beready at the end of the monthand, at the moment, we are not able to make any order (indeed we are changing oursystemof prurchase).At the end of the month however, everything will be OK and we will be able to order again. I could then regularize the order if you are agree.
    Let me know your decision, hoping you will be able to send mean aliquotas soon as possible.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 16 2012

    Answer

    Thank you for contacting Abcam.

    We are unable to send you any free/trail sizes of our antibodies. When your ordering system is reorganized at the end of the month, you will be able to order the antibody and if it is in stock then it will be shipped overnight for next day delivery. Therefore you will still be able to use the antibody on your fresh samples.

    Please let me know if there is anything else I can help you with.

    Read More

    Abcam Scientific Support

    Answered on Jan 16 2012

    Question

    Multiple bands in addition to specific band,using human primary blood samples.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 06 2012

    Answer

    Thank you for your call today and for letting us know about the trouble with ab37185. As we discussed, please let me know if the suggestions that we discussed don't improve the results, and I will be happy to help you further. If you have any questions or if there is anything else that we can do for you, don't hesitate to let me know. Have a great weekend!

    Read More

    Abcam Scientific Support

    Answered on Jan 06 2012

    Question

    Called because he did not receive the email back from technical support when he sent in the questionnaire.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 04 2006

    Answer

    Thank you for your enquiry. As discussed on the phone, here is the email that was sent in response to your questionnaire from Kate Hayes on December 1st, 2006. Reply from Abcam to your enquiry regarding ab37185 Dear Ilangovan, In response to your enquiry (ref: 958900): BATCH NUMBER 227973 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE total protein from mouse brain PRIMARY ANTIBODY Abcam, mouse, two different concentrations, one at 0.5 ug/ml and 2.0 ug/ml was used. Incubated for over night at 4 deg C. Three times washed with TBS plus 0.1% tween 20 each for five minujtes DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED no positive and negative control was used ANTIBODY STORAGE CONDITIONS -20 deg Celsius SAMPLE PREPARATION tissue was homogenized in RIPA buffer supplemented with protease inhibitors AMOUNT OF PROTEIN LOADED 30 microgram ELECTROPHORESIS/GEL CONDITIONS Reducing, 10% SDS-PAGE TRANSFER AND BLOCKING CONDITIONS Transfer buffer 5.8 g Tris, 2.9 g Glycine, 0.37 g SDS and 200 ml methanol adjusted to 1 liter with deionized water, semi-dry transfer for one hour at 25 V, blocked with 5% non-fat dry milk solution SECONDARY ANTIBODY Invitrogen, goat-anti-rabbit, 1:2000, one hour, three times washed with TBS plus tween 20 each for five min. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? none of the steps altered ADDITIONAL NOTES in xCT knock out samples, the protein is appearing with this antibody. We have an answer: Thank you for taking the time to complete our questionaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody (ab37185). I would like to offer some suggestions to optimise the results. I would also appreciate if you can confirm some details of the procedure. 1. In order to increase the signal,I can suggest increasing the antibody concentration. Dilutions on the datasheet are a guideline only. You can try 1:200 dilution. 2. Can you confirm you have reduced and denatured the samples in SDS and mercaptoethanol? This will ensure the protein is in the correct conformation to run at the correct molecular weight and be detected by the antibody. 3. Can you confirm if you have checked protein transfer onto the membrane with Poncea red? 4. How long was the blocking incubation? To ensure the membrane is not overblocked, I suggest this can be reduced. Try 1 hour, with just 3% blocking agent. BSA often gives better results than milk. 5. If you are obtaining faint bands at the correct size for xCT in the knockout samples, it could be that the knockout has not been efficient. Has this been checked, for example using PCR to detect if the xCT gene is still present? I would like to be able to suggest a negative control, but the xCT protein is ubiquitously expressed. I hope this information is helpful. Should the suggestions not improve the results, please do not hesitate to contact me again so I can investigate further. Best regards Kate Kate Hayes Technical Support Assistant Abcam plc https://www.abcam.com

    Read More

    Abcam Scientific Support

    Answered on Dec 04 2006

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